CHAPTER 39
Muscles
M uscles use actin and myosin to generate powerful,
unidirectional movements (Fig. 39.1). The molecular
strategies are specialized versions of those used by
other cells to produce contractions, to adhere to each
other and the extracellular matrix, and to control their
activity.
Vertebrates have three types of specialized contractile
cells: smooth muscle, skeletal muscle, and cardiac
muscle. These muscles have much in common, but differ
in their activation mechanisms, arrangement of contrac-
tile filaments, and energy supplies. The nervous system
controls the timing, force, and speed of skeletal muscle
contraction over a wide range. Cardiac muscle generates
its own rhythmic contractions that spread through the
heart in a highly reproducible fashion. Neurotransmit-
ters, acting like hormones, regulate the force and
frequency of heartbeats over a narrow range. Nerves,
hormones, and intrinsic signals control the activity of
smooth muscles, which contract slowly but maintain
tension very efficiently. This chapter explains the molec-
ular and cellular basis for these distinctive physiological
properties of the three types of muscle.
Skeletal Muscle
Skeletal muscle cells (also called muscle fibers in the
physiological literature) are among the largest cells of
vertebrates. During development, mesenchymal stem
cells give rise to progenitor cells with a single nucleus
called myoblasts. A family of master transcription factors,
including MyoD and myogenin, coordinates the expres-
sion of specialized muscle proteins. As they differentiate,
myoblasts fuse and elongate to form muscle cells with
multiple nuclei and lengths ranging from millimeters to
tens of centimeters. The number of muscle cells is deter-
mined genetically and is relatively stable throughout life
even as the size of the cells varies with the level of exer-
cise and nutrition.
A basal lamina (see Fig. 29.17C) surrounds and sup-
ports each muscle cell. At the ends of each cell, actin
thin filaments are anchored to the plasma membrane at
myotendinous junctions, which are similar to adherens
junctions (see Fig. 31.8). There, integrins spanning the
membrane link actin filaments to the basal lamina and to
collagen fibrils of tendons. These physical connections
transmit contractile force to the skeleton.
Mature muscles harbor small numbers of long-lived
stem cells (see Fig. 41.14) with the potential to repair
damage. They are called satellite cells because they are
located inside the basal lamina next to the muscle cells.
Some of these cells are capable of both self-renewal and,
when the muscle is injured, producing progeny that can
differentiate into myoblasts that can fuse with each other
or existing muscle cells to repopulate the muscle. These
features have made satellite cells a focus of research to
treat degenerative diseases of muscle.
Organization of the Actomyosin Apparatus
Skeletal muscle cells are optimized for rapid, forceful
contractions. Accordingly, they have a massive concen-
tration of highly ordered contractile units composed of
actin, myosin, and associated proteins (Fig. 39.2). Actin
and myosin filaments are organized into sarcomeres,
aligned contractile units that give the cells a striped
appearance in the microscope. For this reason, they are
called striated muscles. Myosin uses adenosine triphos-
phate (ATP) hydrolysis to power contraction, which
results from myosin-powered sliding of actin-based thin
filaments past myosin-containing thick filaments. Speed
of contraction is achieved by linking many sarcomeres
in series. Power (force) is achieved by linking multiple
sarcomeres in parallel. Nerve impulses stimulate a tran-
sient rise in cytoplasmic calcium that activates the con-
tractile proteins.
Interdigitation of thick, bipolar, myosin filaments and
thin actin filaments in the sarcomeres of living muscle
671
672 SECTION IX n Cytoskeleton and Cellular Motility
A. Skeletal muscle
B. Cardiac muscle
C. Smooth muscle
FIGURE 39.1 LIGHT MICROGRAPHS AND INTERPRETIVE
DRAWINGS OF HISTOLOGIC SECTIONS OF SKELETAL,
CARDIAC, AND SMOOTH MUSCLES. A, Skeletal muscle cells are
shaped like cylinders and may be up to 50cm long. Multiple nuclei
are located at the periphery near the plasma membrane. Striations
are seen in the inset, a longitudinal section at high magnification.
B, Cardiac muscle cells are striated and have one or two nuclei.
Adhesive junctions called intercalated disks (bright pink vertical bars in
the longitudinal section, top left arrows) bind these short cells together
end to end. C, Smooth muscle cells are spindle shaped with homo-
geneous cytoplasm and single nuclei.
cells is so precise (Fig. 39.3) that it yields an X-ray dif-
fraction pattern (see Fig. 39.11) revealing the spacing of
the filaments and the helical repeats of their subunits to
a resolution of about 3nm. Z disks at both ends of the
sarcomere anchor the barbed ends of the actin filaments,
so their pointed ends are near the center of the sarco-
mere. Myosin heads project from the surface of thick
filaments, whereas their tails form the filament back-
bone. Thick and thin filaments overlap, with the
Section of
sarcomere
Myofibril
Muscle
cell
Muscle
FIGURE 39.2 CONTRACTILE APPARATUS OF STRIATED
MUSCLES. The contractile unit is the sarcomere, an interdigitating
array of thick and thin filaments. Sarcomeres are arranged end to end
into long, rod-shaped myofibrils that run the length of the cell. Mito-
chondria and smooth endoplasmic reticulum separate myofibrils,
which can readily be isolated for functional and biochemical studies.
myosin heads only a few nanometers away from adjacent
actin filaments. The alignment and interdigitation of the
filaments facilitate the sliding interactions required to
produce contraction.
An important, simplifying architectural feature is that
sarcomeres are symmetrical about their middles (Fig.
39.3). Consequently, the polarity of myosin relative to
the actin filaments is the same in both halves of the sar-
comere, allowing the same force-generating mechanism
to work at both ends of the bipolar myosin filaments.
Sarcomeres are organized end to end into long, rod-
shaped assemblies called myofibrils (Fig. 39.2) that
retain their contractility even after isolation from muscle.
Thin Filaments
Thin filaments are a polymer of actin with tightly bound
regulatory proteins troponin and tropomyosin (Fig.
39.4). When the cytoplasmic Ca2+ concentration is low,
troponin and tropomyosin inhibit the actin-activated
adenosine triphosphatase (ATPase) of myosin. Tropo-
myosin, a 40-nm long coiled-coil of two -helical poly-
peptides (see Fig. 3.10), binds laterally to seven

A A
C Barbed
B D
A band I band
Z disk M line
Myosin Actin Myosin
E polarity polarity polarity
FIGURE 39.3 ELECTRON MICROGRAPHS AND DRAWINGS
OF SARCOMERES. A, Longitudinal thin section showing the array of
thin filaments anchored to Z disks and overlapping bipolar thick fila-
ments crosslinked in the middle at the M line. B, Longitudinal freeze-
fractured, etched, and shadowed sarcomere showing myosin
cross-bridges attached to thin filaments near the bare zone in the
center (right) of a sarcomere. CD, Cross-sections of insect flight
muscle and vertebrate skeletal muscle showing the double hexagonal
arrays of thick and thin filaments. E, Drawings indicating the polarity
of the thick and thin filaments. (A and C, Courtesy H.E. Huxley,
Brandeis University, Waltham, MA. B and D, Courtesy J. Heuser,
Washington University, St. Louis, MO.)
contiguous actin subunits as well as head to tail to neigh-
boring tropomyosins, forming a continuous strand along
the whole thin filament. Troponin (TN) consists of three
different subunits called TNC, TNI, and TNT (see Table
39.1). TNT anchors one troponin complex to each tropo-
myosin coiled-coil. TNC is a dumbbell-shaped protein
with four EF-hand motifs to bind divalent cations similar
to calmodulin (see Fig. 3.12 and Chapter 26). In resting
muscle, the C-terminal globular domain of TNC binds
two Mg2+ ions and an -helix of TNI, while the low-
affinity sites in the N-terminal globular domain of TNC
CHAPTER 39 n Muscles 673
Tropomyosin Tropomodulin
Troponin
Pointed
B
C C
N
N
C Troponin C alone Troponin C on TNI peptide
FIGURE 39.4 THIN FILAMENT STRUCTURE. A, Three-
dimensional reconstruction from electron micrographs of a thin fila-
ment from vertebrate skeletal muscle showing actin and the position
of tropomyosin in relaxed muscle. B, Drawing of a model of a thin fila-
ment from active muscle based on reconstructions of electron micro-
graphs and crystal structures of troponin and tropomodulin. Each
troponintropomyosin unit is associated with seven actin subunits.
C, Ribbon diagrams of the atomic structures of troponin C, free and
bound to a troponin I peptide. Two divalent cation-binding EF hands
are found at each end, separated by a long -helix. In cells, two high-
affinity sites at the C-terminal end are permanently occupied with Mg2+.
Two low-affinity sites at the other end are unoccupied in relaxed muscle
but bind Ca2+ when muscle is activated. (A, Based on Protein Data
Bank [www.rcsb.org] file 1AX2, created by Roberto Dominguez,
University of Pennsylvania.)
are empty. Ca2+ binding to the low-affinity sites (two in
fast skeletal muscle; one in slow muscle) during muscle
activation exposes a new binding site for TNI. The result-
ing conformational change in TNI allows tropomyosin to
move away from the myosin-binding sites on the actin
filament.
A protein meshwork in the Z disk anchors the barbed
end of each thin filament (Fig. 39.5). Some crosslinks
between actin filaments consist of -actinin, a short rod
with actin-binding sites on each end (see Fig. 33.17). At
least a half dozen structural proteins stabilize the Z disk
through interactions with -actinin, actin, and titin.
Some of these proteins also have signaling functions.
Proteins cap both ends of thin filaments. Cap-Z, the
muscle isoform of capping protein (see Fig. 33.15), binds
the barbed ends of thin filaments with high affinity, limit-
ing actin subunit addition or loss. Tropomodulin asso-
ciates with both tropomyosin and actin to cap and
stabilize the pointed end of the thin filament (Fig. 39.4B).
Tropomyosin and a gigantic filamentous protein,
nebulin, stabilize thin filaments laterally. Nebulin con-
sists of 185 imperfect repeats of a 35-amino-acid motif
674 SECTION IX n Cytoskeleton and Cellular Motility
A. Longitudinal section B. Cross section
Z disk
C
FIGURE 39.5 Z DISK STRUCTURE. AB, Electron micrographs
of thin sections perpendicular to and in the plane of the Z disk.
C, Three-dimensional reconstruction, based on electron micrographs
of the Z disk, showing the network of protein crosslinks that anchor
the barbed ends of the yellow actin filaments. (Courtesy J. Deatherage,
National Institutes of Health, Bethesda, MD and modified from Cheng
NQ, Deatherage JF. Three dimensional reconstruction of the Z disk of
sectioned bee flight muscle. J Cell Biol. 1989;108:17611774.)
that interact with each actin subunit, tropomyosin, and
troponin along the length of thin filaments. Interactions
with tropomodulin and Z disk proteins anchor nebulin
at the two ends of the thin filament. These interactions
influence the length of thin filaments, but nebulin does
not act simply as a molecular ruler.
Thick Filaments
The self-assembly of myosin II (see Fig. 5.7) establishes
the bipolar architecture of striated muscle thick fila-
ments (Fig. 39.6). Some features of thick filaments are
invariant, such as a superhelical backbone consisting of
myosin tails, a surface array of myosin heads, the 14.3-nm
stagger between rows of heads, and a central bare zone
formed by antiparallel packing of tails. Phosphorylation
of the myosin regulatory light chains favors release of
the myosin heads from the surface of the filament in
preparation for engagement with actin. Filaments may
A
B C
D
M line
E
Bare zone
FIGURE 39.6 STRUCTURE OF BIPOLAR THICK FILAMENTS.
A, Electron micrograph of a thick filament isolated directly from skeletal
muscle and prepared by negative staining. A single myosin molecule
is shown at the same magnification at the lower left. The myosin tails
form the backbone of the thick filament and allow the two myosin
heads to swing out from the side (see the enlarged inset on the
right). B, Reconstruction from cryoelectron micrographs of part of a
tarantula skeletal muscle thick filament with the bare zone (not shown)
to the right. The tails form the backbone of the filament and the myosin
heads are folded back toward the bare zone. Space-filling models of
the two heads of one myosin molecule are superimposed on the
reconstruction. The catalytic domains are green and blue; the light
chains are pink, orange, yellow, and tan. C, Cross section of vertebrate
skeletal muscle showing the double hexagonal arrays of thick and thin
filaments. D, Electron micrograph of a highly stretched sarcomere with
the M line in the middle. Note the nine faint stripes of myosin-binding
protein C along both halves of the think filaments. E, Drawing of protein
links between thick filaments in the M line. (A, Courtesy John Trinick,
University of Bristol, United Kingdom. For reference, see Knight P,
Trinick J. Structure of the myosin projections on native thick filaments
from vertebrate skeletal muscle. J Mol Biol. 1984;177:461482.
B, Courtesy J. Woodhead and R. Craig, University of Massachusetts
Medical School, Worcester, MA. For reference, see Woodhead JL,
Zhao FQ, Craig R, et al. Atomic model of a myosin filament in the
relaxed state. Nature. 2005;436:11951199. C, Courtesy J. Heuser,
Washington University, St. Louis, MO. D, Courtesy H.E. Huxley,
Brandeis University, Waltham, MA.)
vary in length, diameter, and organization of the helical
array of heads in various species. Invertebrate thick fila-
ments have a core of paramyosin, a second coiled-coil
protein, which is not found in vertebrates.
Accessory proteins stabilize thick filaments in striated
muscles (Table 39.1). Myosin-binding protein C, with
 CHAPTER 39 n Muscles 675

its fibronectin III and immunoglobulin domains, forms

nine stripes along both halves of the thick filament (Fig.
39.6D) and also interacts with thin filaments. It fine- PEVK
tunes the interactions of myosin heads with both fila-
ments, maintaining the relaxed state but also favoring
crossbridge formation when the muscle is activated. The
M line in the center of the sarcomere of most types
of muscle (Fig. 39.6D) is a three-dimensional array of
protein crosslinks. These elastic crosslinks maintain the
precise registration of thick filaments but also allow the
filaments to move apart as the sarcomere shortens while
maintaining a constant volume. At least three structural Titin
proteins and the enzyme MM-creatine phosphokinase
(which transfers phosphate from creatine-phosphate to PEVK
adenosine diphosphate [ADP]) are located in the M line. Ig domains Sarcomere length = 2.4 m
PEVK
unfolds
Titin Filaments
Titin, the largest protein encoded by the human genome, Chain of Ig domains Stretched to 3.2 m
stretched (physiological stretch length)
forms a third array of filaments lying parallel to the thin
and thick filaments and connecting the Z disk to the
thick filaments and the M line (Fig. 39.7). Three titins Ig domains unfold Stretched to 3.6 m
flank each half thick filament. Although titin is the third
most abundant protein in muscle, it is hard to preserve FIGURE 39.7 TITIN FILAMENTS. Top panel, Electron micro-
for electron microscopy, so these diaphanous filaments graphs of single, isolated titin molecules prepared by heavy metal
shadowing. Titin molecules are long enough to extend from the Z disk
escaped notice for years. Each filament is a single poly-
to the M line. Middle panel, Drawing of a sarcomere, to the same scale
peptide named after a mythological giant owing to its as the electron micrograph, with the thick filaments removed from the
remarkable size: several splice isoforms consist of 27,000 bottom half to illustrate how titin molecules anchor thick filaments to
to 33,000 amino acids folded into a linear array of up to the Z disk and extend to the M line. Bottom panel, Drawing illustrating
300 fibronectin III and immunoglobulin (Ig) domains a model for the elasticity of titin. Modest stretches within the physio-
logical range reversibly extend the chain of immunoglobulin (Ig)
measuring more than 1.2m long.
domains in the I-band and the PEVK domain. Extreme extension can
The elasticity of titin molecules provides passive resis- unfold Ig domains. (Modified from Reif M, Gautel M, Oesterhelt F, etal.
tance to stretching of relaxed muscle. Titin connections Reversible unfolding of individual titin immunoglobulin domains by
to the Z disk and thick filaments provide physical continu- AFM. Science. 1997;276:10901092. For reference, see Leake MC,
ity from one sarcomere to the next and keep the thick fila- Wilson D, Gautel M, Simmons RM. The elasticity of single titin
molecules using a two-bead optical tweezers assay. Biophys J.
ments centered in the sarcomere during contraction.
2004;87:11121135.)
Differential splicing creates titin isoforms that differ in
stiffness for various types of muscle. If titin molecules are
broken experimentally, thick filaments slide out of regis-
ter toward one Z disk during contraction. Two features
provide the elasticity during short (~0.3m per titin),
physiological stretching (Fig. 39.7). The irregular chain of
Ig domains in the I band straightens out, and a segment Intermediate
of the polypeptide rich in proline, glutamic acid, valine, filaments
and lysine (the PEVK domain) partly unfolds. This
decreases entropy and provides the energy for elastic
recoil (an entropic spring; see also Fig. 29.11). Extreme
stretching unfolds Ig domains one by one. Titin not only
responds to phosphorylation but also binds signaling
proteins that influence the performance of muscle in the
short and long term.
M line
Z disk
Intermediate Filaments
FIGURE 39.8 DESMIN INTERMEDIATE FILAMENTS IN SKEL-
Desmin intermediate filaments (see Chapter 35) help
ETAL MUSCLE. Desmin filaments connect Z disks laterally to each
align the sarcomeres laterally (Fig. 39.8) by linking each other and to the plasma membrane at specializations called costa-
Z disk to its neighbors and to specialized attachment meres. (Modified from Lazarides E. Intermediate filaments as mechani-
sites on the plasma membrane called costameres. In cal integrators of cellular space. Nature. 1980;283:249256.)
676 SECTION IX n Cytoskeleton and Cellular Motility
A. Relaxed and stretched
B. Contracted
Crossbridges
C. Rigor showing crossbridges
FIGURE 39.9 SLIDING FILAMENTS. Electron micrographs and
interpretive drawings of longitudinal sections of a sarcomere from a
relaxed muscle (A) and a contracted skeletal muscle (B). The lengths
of the thin and thick filaments are constant as the sarcomere shortens,
demonstrating that the filaments slide past each other during contrac-
tion. C, Crossbridges between thick and thin filaments from a muscle
in rigor. (Micrographs courtesy H.E. Huxley, Brandeis University,
Waltham, MA.)
addition to desmin, costameres contain clathrin plus
several cytoskeletal proteins (vinculin, talin, spectrin,
and ankyrin) found in focal contacts and adherens junc-
tions of nonmuscle cells (see Figs. 30.11 and 31.8).
Desmin mutations in humans cause disorganization of
myofibrils, resulting in generalized muscle failure.
Molecular Basis of Skeletal Muscle Contraction
Sliding Filament Mechanism
The key to understanding muscle contraction was the
discovery that thick and thin filaments maintain constant
lengths and slide past each other as sarcomeres (and the
muscle) shorten (Fig. 39.9). About the same time, it was
discovered that crossbridges (now recognized to be
myosin heads) can connect actin and myosin filaments,
and that tension produced during contraction is propor-
tional to the overlap of actin and myosin filaments (Fig.
39.10). Supported by biochemical and ultrastructural
evidence for actinmyosin interaction, these pioneering
observations led to the theory that crossbridges between
the thick and thin filaments produce the force for con-
traction. Sixty years of research on crossbridges have
yielded a detailed picture of the chemistry and molecular
mechanics underlying the force-producing reactions. A
review of the steps of the actomyosinATPase cycle (see
Fig. 36.5) is helpful in understanding the contraction
mechanism. Three different physiological states reveal
information about crossbridge mechanisms.
Relaxed. One extreme is relaxed muscle. When the
concentration of cytoplasmic Ca2+ is low, tropomyosin
b
a
Force
Force
c
0 0
A Sarcomere length C Velocity
a
c
B
FIGURE 39.10 PHYSIOLOGICAL PROPERTIES OF SKELETAL
MUSCLE. A, Dependence of maximum tension on the length of
the sarcomeres. B, Interpretive drawings. Each relates to a point
on A. C, Relationship of force and velocity during muscle contraction.
(A, For reference, see Gordon AM, Huxley AF, Julian F. The variation
in isometric tension with sarcomere length in vertebrate muscle fibres.
J Physiol. 1964;171:28P30P. C, From Ruch TC, Patton HD (eds).
Physiology and Biophysics, 19th ed. Philadelphia: WB Saunders;
1965.)
and troponin inhibit the interaction of myosin heads
with actin filaments, resulting in few myosin heads
being bound. Lacking long-lived physical connections
between the filaments, muscle offers little resistance
to passive stretching. X-ray diffraction (Fig. 39.11)
shows that the myosin heads (with bound ATP or
ADP and phosphate) are closely associated with the
backbone of thick filaments and arranged in a helical
array determined by the thick filament structure
(Fig. 39.6B).
Rigor. The other extreme occurs after death. Depletion
of ATP allows all myosin heads to bind tightly to
actin filaments (Figs. 39.3B and 39.9C). By X-ray dif-
fraction, the myosin heads bound to actin filaments
contribute to the strength of the reflections from
the actin filament helix. Strong physical connections
between the filaments prevent stretching, making
the muscle stiff (hence the term rigor mortis). This
extreme condition illustrates what happens structur-
ally and mechanically when all the crossbridges
engage actin filaments.
Contracting. The most interesting, but most compli-
cated, state is actively contracting muscle. Myosin
heads walk along actin filaments toward their
barbed ends, pulling Z disks toward the center of
the sarcomere. Thousands of sarcomeres shorten in
series, causing the whole muscle to shorten. ATP is
consumed and force is produced. The thick filament
 CHAPTER 39 n Muscles 677

A B. Relaxed C. Contracting

Myosin
head
helix

Actin Actin
helix helix

Heads along thick filament Dancing heads

FIGURE 39.11 CROSSBRIDGE DYNAMICS REVEALED BY X-RAY DIFFRACTION PATTERNS OF WHOLE MUSCLE. A, Electron
micrograph showing the orientation of the muscle in the x-ray beam in B and C. BC, Fiber diffraction patterns from relaxed and contracting
skeletal muscles with interpretive drawings of crossbridges in each state. Reflections from myosin heads arranged on the thick filament helix are
strong in relaxed muscle. Reflections from the actin helix are stronger than the thick filament helix in contraction. The myosin and actin reflections
are each labeled in only one of four equivalent quadrants. During contraction, a few myosin heads attach transiently to actin, increasing the strength
of the actin helix reflections, but most are disordered. (Micrograph and x-ray patterns courtesy H.E. Huxley, Brandeis University, Waltham, MA.)

helical pattern is very weak by X-ray diffraction
(Fig. 39.11C). Actin reflections are stronger than
relaxed muscle but not as strong as rigor. Disordered
myosin heads are distributed between the thick and
thin filaments as each one dances asynchronously on
and off of actin filaments.
Most myosin heads in contracting muscle have bound
ATP or ADP-Pi (adenosine diphosphateinorganic phos-
phate), allowing them to exchange rapidly among the
four weakly bound states illustrated in Fig. 36.5. During
some of the transient interactions of myosinADP-Pi with
actin, phosphate dissociates from myosin, and the light-
chain domain rapidly reorients (see Figs. 36.4 and 36.5).
This stretches elastic elements in the myosin heads and
both thick and thin filaments. Energy in these elastic
elements can be used over a period of milliseconds to
displace the actin filament relative to the crossbridge and
contract the muscle. When ADP dissociates from the
actinmyosinADP intermediate, ATP rapidly binds to
the actinmyosin complex, dissociating the crossbridge
and starting a new ATPase cycle.
Relationship of Crossbridge Behavior to
the Mechanical Properties of Muscle
Normally, each sarcomere shortens less than 1m.
However, the whole muscle shortens macroscopically,
because it has thousands of sarcomeres in series. For
example, a human biceps muscle 20cm long has approx-
imately 80,000 sarcomeres in series from end to end.
When each contracts 0.25m, the muscle shortens
2cm. Because the system maintains a constant volume,
each sarcomere and the whole muscle increase in diam-
eter as they shorten. Although the individual filaments
slide past each other relatively slowly (about 2m s1 in
both halves of each sarcomere), muscles contract rapidly
because the motion of each sarcomere in the series is
added together. In our example, without resistance, the
biceps contracts approximately 3cm in 100ms during
which most crossbridges pull just once.
Crossbridge behavior explains why the velocity of
muscle contractions of an active muscle depends on the
external load (Fig. 39.10). When opposed by no load the
molecular motion stored in elastic elements of each
crossbridge is largely converted into movement of actin
filaments relative to myosin filaments and contraction
velocity is maximal. Under these conditions, the fila-
ments in muscle slide past each other at a rate of about
5m s1, the same speed observed for free actin fila-
ments moving over myosin heads in vitro (see Fig. 36.6).
For this rapid sliding to occur, myosin heads that do not
produce force must not impede movement. If bound
tightly to actin, they would interfere mechanically with
rapid sliding. This is avoided by the rapid equilibrium of
the myosin intermediates between being bound to actin
and being free. Transient interactions of myosin heads
678 SECTION IX n Cytoskeleton and Cellular Motility
bound to ATP or ADP-Pi with actin do not produce force
or retard sliding driven by force-producing crossbridges.
Muscle produces maximum force when the contrac-
tion rate is zero (Fig. 39.10). The conformational change
in the myosin head stretches elastic elements in the
crossbridge, but the force cannot overcome the resis-
tance from the load on the muscle. Consequently, the
filaments do not slide, and energy stored in each stretched
elastic element is lost as heat when the crossbridge dis-
sociates at the end of the ATPase cycle. The maximum
force depends on the numbers of sarcomeres in parallel,
that is, the cross-sectional area of the muscle. This
explains why muscles respond to strengthening exer-
cises by growing in diameter.
Regulation of Skeletal Muscle Contraction
Although skeletal muscle cells have only two states
inactive (relaxed) or active (contracting)skeletal
muscles produce a wide range of contractions, varying
from slow and delicate to rapid and forceful. These
graded contractions are achieved by varying the
number of muscle cells activated by voluntary or reflex
signals from the nervous system (Fig. 39.12).
Control of Skeletal Muscle by Motor Neurons
Neural stimuli that activate skeletal muscles arise in two
ways (Fig. 39.12). In organisms with well-developed
central nervous systems, most neural signals that activate
skeletal muscles result from conscious decisions, provid-
ing voluntary control over skeletal muscles. Other signals
Sensory neuron
Brain
Dorsal root
Ventral root Sensory nerve
Spinal Motor neuron Motor nerve
cord
Neuromuscular
junction
Skeletal Sensory
muscle cell muscle
spindle cell
FIGURE 39.12 INNERVATION OF SKELETAL MUSCLE. Motor
neurons in the spinal cord stimulate one or (usually) more skeletal
muscle cells. Two neural pathways control motor neurons. Some
stimuli come from neurons in higher centers of the brain. This pathway
provides voluntary control over muscle contraction. Other stimuli come
through local reflex circuits from sensory detectors, including muscle
spindle cells. These signals help coordinate muscle contraction in
response to changing forces on and within the muscle.
result from reflex responses to stimulation of sensory
nerves. Specialized muscle cells innervated with both
motor and sensory nerves function as stretch receptors,
relaying information about length and tension back to
the spinal cord, where reflexes coordinate the motor
neuron output. Neural inputs from both sources con-
verge on motor neurons located in the brainstem and
spinal cord of vertebrates. Axons of these motor neurons
branch in a muscle to contact one or more muscle cells.
A motor neuron together with its target muscle cells
forms a motor unit. In the most precisely controlled
muscles, such as the extraocular muscles in the eye,
some motor neurons innervate single muscle cells.
The contractile activity of a muscle is graded in terms
of the speed and force of the contraction, so individual
muscles can produce both delicate and powerful move-
ments. Nerve stimulation determines the contractile
force in two ways: (a) The number of active motor units
determines how many muscle cells produce force, and
(b) the rate of stimulation adjusts the force produced
by active cells. Every time a muscle cell is stimulated, all
the sarcomeres are activated, but the force that they
produce increases as the rate of stimulation increases,
up to a maximum of approximately 200 stimuli per
second. The shortening velocity of an active muscle
depends on the ratio between force produced and the
resistance (Fig. 39.10C). If a large force or high velocity
of contraction is required, many motor units are called
into action. To sustain contraction, motor nerves fire
repeatedly. By varying the number of active cells in a
muscle and the rate of stimulation, the nervous system
sets the force required for a particular movement.
Synaptic Transmission at Neuromuscular Junctions
The terminal branch of each motor neuron axon forms
a large synapse called the motor end plate or neuro-
muscular junction on the muscle surface (see Fig.
17.9). These nerve endings are filled with synaptic vesi-
cles containing the neurotransmitter acetylcholine.
Arrival of an action potential at the nerve terminal
stimulates fusion of synaptic vesicles with the nerve
plasma membrane, releasing acetylcholine into the cleft
between nerve and muscle. In less than a millisecond,
acetylcholine diffuses across the extracellular space
and binds to acetylcholine receptors concentrated in
the adjacent muscle plasma membrane. Acetylcholine
binding opens the receptor cation channel, initiating a
new action potential that spreads over the muscle cell
plasma membrane and intracellularly into the T tubules.
Coupling Action Potentials to Contraction
The plasma membrane of skeletal muscle cells, like that
of nerve cells, is excitable (see Fig. 17.6) but, unlike
that in nerves, it invaginates deeply to form T tubules
that run across the entire cell (Fig. 39.13). Depending
on the species and type of striated muscle (skeletal
 CHAPTER 39 n Muscles 679

versus cardiac), T tubules may be located either at the

A Plasma Entrance to level of the Z disks or at the thick filament ends. Inside
membrane T tubule
the muscle cell, T tubules interact extensively with the
smooth endoplasmic reticulum (SER) surrounding each
myofibril. Historically, this SER was called sarcoplasmic
reticulum. Terminal cisternae of SER are closely associ-
ated with T tubules at foot processes that can be
visualized by electron microscopy. Together, T tubules
and SER constitute a signal-transducing apparatus that
converts depolarizations of the plasma membrane
T tubule
into a spike of cytoplasmic Ca2+ to trigger contraction
Smooth
endoplasmic (Fig. 39.14).
reticulum An action potential moving through a T tubule trig-
gers the release of Ca2+ from SER into the cytoplasm (Fig.
B 39.14). Ca2+ binding to troponin allows myosin to inter-
act with the thin filament, initiating contraction. This
C. Skeletal muscle signal transduction process is called excitation
T tubule contraction coupling. Three transmembrane proteins
located in the T tubule and the terminal cisternae of
the SER cooperate to generate the transient Ca2+ signal
(Fig. 39.14).
1. A voltage-sensitive calcium channel (see Chapter 16)
senses action potentials in the T tubule. These chan-
nels are called dihydropyridine (DHP) receptors,
owing to their affinity for this class of drugs. The
actual Ca2+ channel of DHP receptors is not essential
for skeletal muscle contraction, as external Ca2+ is not
required for contraction in the short term.
Smooth ER
2. Ca2+ release channels (see Fig. 26.13), concentrated
in the terminal cisternae of SER, release Ca2+ into the
D. Cardiac muscle cytoplasm. A drug called ryanodine binds these chan-
Smooth ER T tubule
nels and inhibits Ca2+ release. Every second ryano-
dine receptor is connected to cytoplasmic loops
of four DHP receptors, forming bridges called foot
processes between the T tubule and the endoplasmic
reticulum (Fig. 39.13B).
3. The P-type calcium-ATPase (see Fig. 14.7) actively
pumps Ca2+ from cytoplasm into the endoplasmic
reticulum against a concentration gradient greater
than 104. Inside the SER several low-affinity, high-
capacity Ca2+-binding proteins buffer the millimolar
concentration of Ca2+. For example, numerous car-
boxyl groups on the surface of calsequestrin bind
Ca2+ with a millimolar Kd. This rapidly reversible reac-
FIGURE 39.13 PLASMA MEMBRANE SPECIALIZATIONS OF
STRIATED MUSCLES. AB, Electron micrographs of thin sections of tion increases the Ca2+ storage capacity of endoplas-
fish skeletal muscle showing plasma membrane invaginations called T mic reticulum without sacrificing the speed of Ca2+
tubules, which cross the whole muscle cell and associate closely with release. Accessory subunits anchor calsequestrin to
smooth endoplasmic reticulum (ER). The complex of a T tubule with Ca2+ release channels, ensuring a local supply of Ca2+
smooth ER on both sides is called a triad. A, A longitudinal section of
for release into cytoplasm when muscle is activated.
two T tubules. B, A cross section of a T tubule flanked on two sides
by smooth ER. Foot processes, consisting of voltage-sensitive calcium An action potential in a T tubule results in a transient
channels in the T tubule paired with calcium release channels in the rise in cytoplasmic Ca2+, from 0.1M to about 2M
ER, connect the T tubule to the smooth ER (see Fig. 39.14 for molecu- (Fig. 39.15), as follows. The action potential causes a
lar details). CD, The three-dimensional arrangement of T tubules and short-lived conformational change in the DHP receptors
smooth ER relative to the sarcomeres in skeletal and cardiac muscle.
that is transmitted mechanically to associated ryanodine
(AB, Courtesy C. Franzini-Armstrong and K. Porter, University of
Pennsylvania.) receptor Ca2+ release channels. Many Ca2+ channels
open transiently, allowing Ca2+ to diffuse down the steep
680 SECTION IX n Cytoskeleton and Cellular Motility

A. Skeletal muscle MYOFIBRIL B. Cardiac muscle

V V

Ca2+
SER T TUBULE SER Ca2+
Ca2+
Voltage-
Ca2+ sensitive Ca2+ Ca2+
channel
Ca2+
Ca2+

Ca2+ Ca2+
Ca2+ Ca2+

ADP
ATP ATP
ADP Ca2+ Ca2+

Ca2+ Ca2+ Ca2+ Ca2+

Ca2+ Ca2+ Ca2+ Ca2+ Ca2+ Ca2+ Ca2+ Ca2+

Ca2+
Ca2+ Ca2+ Ca2+

Ca2+
Ca2+

Ca2+ Ca2+
Ca2+ Ca2+ Ca2+

ADP
ADP
ATP
Ca2+ ATP Ca2+
V V Ca2+
Ca2+

FIGURE 39.14 MECHANISM OF CALCIUM RELEASE IN SKELETAL AND CARDIAC MUSCLES. Both muscle types use voltage-sensitive
calcium channels in the T tubule membranes and calcium release channels in the smooth endoplasmic reticulum (SER). A, Direct coupling in
skeletal muscle. An action potential in the T tubule (V) activates the voltage sensor (turning from gray to blue). This direct contact opens the
calcium release channel (turning from gray to pink). Cytoplasmic Ca2+ levels rise only briefly because calciumATPase (adenosine triphosphatase)
pumps Ca2+ back into the lumen of the SER. B, Calcium-induced Ca2+ release in cardiac muscle. An action potential opens the voltage-sensitive
Ca2+ channel in the T tubule, releasing Ca2+ into the cytoplasm. This Ca2+ opens the calcium release channel in the SER. ADP, adenosine diphos-
phate; ATP, adenosine triphosphate.

concentration gradient from the SER lumen to the cyto-
plasm. Physical connections between ryanodine recep-
tors may spread their activation laterally, ensuring
synchronous activation of a patch of channels.
After a single action potential, the free Ca2+ in the cyto-
plasm rises for only a few milliseconds for three reasons.
First, Ca2+ release channels close quickly. Second, cyto-
plasmic Ca2+ binds to troponin C and other proteins.
Third, Ca2+ pumps efficiently transport cytoplasmic Ca2+
back into the lumen of the SER, even before the muscle
develops maximum force. Ca2+ pumps are continuously
active, keeping the cytoplasmic Ca2+ concentration low.
This is why repeated action potentials are required to
prolong the rise in cytoplasmic Ca2+ (Fig. 39.15B).
Transduction of the Calcium Spike Into Contraction
Troponintropomyosin on thin filaments cooperates
with myosin to turn on contraction in response to a Ca2+
spike. At rest, two Ca2+-binding sites of fast skeletal
muscle troponin C are largely unoccupied (owing to
their low affinity for Ca2+ and the low Ca2+ concentra-
tion). As a result, the troponintropomyosin complex
partially blocks the binding site for myosin heads on
actin (Fig. 39.16). This prevents most of the weak-binding

A. Twitch
Stimulus
Force
Ca2+
Force
Ca2+
Milliseconds
B. Tetanus
Stimulus
Force
Force
Ca2+
Ca2+
Milliseconds
FIGURE 39.15 CA2+ TRIGGERS CONTRACTION OF SKELE-
TAL MUSCLE. In these experiments, the Ca2+-sensitive protein
aequorin was injected into live muscle cells to provide a signal for the
cytoplasmic Ca2+ concentration. A, Single stimulus. Cytoplasmic Ca2+
concentration increases transiently, followed by a short contraction.
This brief contraction persists after cytoplasmic Ca2+ decreases to the
resting level. B, Multiple stimuli. Each stimulus releases a new pulse
of Ca2+, prolonging the contraction in so-called tetanus. (For reference,
see Ridgway EB, Ashley CC. Calcium transients in single muscle
fibers. Biochem Biophys Res Commun. 1967;29:229234.)
myosin intermediates with ATP or ADP-Pi in the active
site from binding the thin filament. When released into
cytoplasm, Ca2+ binds troponin C, causing a conforma-
tional change that creates a binding site for a helical
region of TNI. This interaction attracts the C-terminus of
TNI away from actin and tropomyosin, allowing a small
shift in the position of tropomyosin on the thin filament.
This shift increases the probability that myosin-ADP-Pi
heads will bind to the thin filament, dissociating their
bound Pi and producing force. Activation is cooperative
for three reasons: Ca2+ must occupy both binding sites
on troponin C, the effects of Ca2+ binding and myosin
binding are transmitted to neighboring tropomyosins
through their end-to-end attachments, and every myosin
that binds accentuates the response. This cooperativity
makes the onoff switch respond very sharply to a rela-
tively small, 10- to 20-fold change in the cytoplasmic Ca2+
concentration. The efficiency of this switch is under-
scored by the fact that the energy consumption of a
muscle cell increases more than 1000-fold when it is
activated. Activation of slow skeletal muscle (Table 39.3)
and cardiac muscle is less cooperative, as their troponin
C has only one Ca2+-binding site.
Note that the muscle produces force well after the
cytoplasmic Ca2+ concentration returns to resting levels
(Fig. 39.15). The Ca2+-sensitive switch is sharp but rela-
tively slow owing to the slow response of thin filaments
CHAPTER 39 n Muscles 681
active
Active
ed
lly
Partia
Relax
A
Myosin-
binding site
Relaxed
Partially active
Active
B
FIGURE 39.16 THIN FILAMENT ACTIVATION MECHANISM.
Reconstructions from electron micrographs showing a short segment
of thin filament (A) and a cross section of a thin filament (B). Ca2+
binding to troponin C partially activates the filament by moving tropo-
myosin away from its lateral position in relaxed muscle, where it over-
laps the myosin-binding site on actin (red). Myosin binding to the
partially activated filament shoves tropomyosin further out of the way
into the active position. (Data from W. Lehman, Boston University, MA.)
to Ca2+ binding. Ca2+ binds troponin C rapidly (milli-
seconds) but dissociates slowly (tens of milliseconds).
Thus, after the Ca2+ spike saturates troponin C and the
thin filament turns on, the muscle remains active even
after free Ca2+ has returned to resting levels. Force
declines slowly as Ca2+ dissociates from troponin C and
returns to the SER without raising the cytoplasmic Ca2+
concentration.
A single action potential produces a short contractile
twitch (Fig. 39.15). Maximum contractile force is pro-
duced by a series of closely spaced action potentials,
leading to a sustained rise in cytoplasmic Ca2+ and pro-
longed activation of actomyosin. The extended contrac-
tion is called tetanus.
Regulation by Myosin Light Chains
The participation of skeletal muscle myosin light chains
in the regulation of contraction varies among species.
The skeletal muscles of mollusks are one extreme. Their
682 SECTION IX n Cytoskeleton and Cellular Motility

TABLE 39.1 Sarcomere Proteins of Vertebrate Striated Muscles

Name Size (kD) Domains Functions Disease Manifestations
Thick Filament
Myosin 2 200 ATPase, coiled-coil Motor, backbone of HCM, HF, arrhythmias
thick filament
Regulatory light chain 2 19 EF-hands Stabilizes lever arm HCM, arrhythmias
Essential light chain 2 18 or 25 EF-hands Stabilizes lever arm HCM, arrhythmias
Myosin-binding protein C 141 Ig, FNIII Stabilizes thick filament HCM, DCM, arrhythmias
Titin 3700 FNIII, IgC2, kinase Elastic connection from DCM, HF, muscular dystrophy
Z disk to M line
M Line
MM-creatine phosphokinase 2 43 Glycolytic enzyme
M protein 165 IgC2, FNIII M-line fast skeletal
muscle
Myomesin (skelemin) 185 IgC2, FNIII Link M-disk to desmin None yet known
Thin Filament
Actin 43 Thin filaments backbone DCM, HF, myopathies
Tropomyosin 2 35 Coiled-coil Blocks myosin binding HCM, DCM, arrhythmias, myopathies
to actin filament
Troponin C 18 EF-hands Calcium-binding DCM likely
Troponin I 21 Inhibitory component HCM, arrhythmias, myopathies
Troponin T 31 Tropomyosin binding HCM, DCM, arrhythmias, myopathies
Tropomodulin 43 Caps actin filament None yet known
pointed end
Nebulin 500900 185 35 residues Binds thin filament Nemaline myopathy
Z Disk
-Actinin 2 100 Actin-binding, Crosslinks thin filaments Focal segmental glomerulosclerosis
spectrin repeats in the Z disk
CapZ 31 + 32 Caps actin filament None yet known
barbed end
Desmin 2 53.5 Intermediate filament Anchors Z disk DCM, myopathy

DCM, dilated cardiomyopathy; EF, calcium-binding helices E and F of calmodulin; FN, fibronectin; HCM, hypertrophic cardiomyopathy; HF, heart failure;
Ig, immunoglobulin; MLCK, myosin light-chain kinase.

myosin light chains bind Ca2+ and provide the main on

TABLE 39.2 Muscle Cell Types
off switch for contraction. When the Ca2+ concentration
is low in resting muscle, no Ca2+ binds to light chains, Physiological Type Myosin Type Mitochondria Fatigue
and the actinmyosin ATPase is off. Ca2+ that is released Fast, white Fast Few Rapid
during activation binds to the light chains, turning on Intermediate Fast Medium Medium
the ATPase and contraction. At the other extreme, the Fast, red Fast Many Slow
light chains of vertebrate skeletal muscle myosin do not Slow, red Slow Many Slow
bind Ca2+, but their phosphorylation modulates contrac-
tile activity by increasing force production at suboptimal
Ca2+ concentrations. Horseshoe crab skeletal muscle
uses a dual system: Ca2+ binding to troponintropomyosin primary transcript (see Fig. 16.6) creates more than 50
on thin filaments and Ca2+-regulated phosphorylation of isoforms of troponin T. Mutations in the genes for
myosin light chains both stimulate contraction. myosin, actin, desmin, nebulin, tropomyosin, troponin-I,
and troponin-T can each cause defects in human skeletal
Specialized Skeletal Muscle Cells muscles (Table 39.1).
All skeletal muscle cells are built on the same principles, Physiological properties, such as the speed of contrac-
but vertebrates actually have several different types of tion and the rate of fatigue, provide criteria for classify-
skeletal muscle cells, each with distinct isoforms of con- ing muscle cells (Table 39.2). The isoforms of myosin
tractile protein and metabolic enzymes. The six myosin (and probably the other contractile proteins) determine
heavy chains and three actin isoforms are coded by dif- the speed of contraction, whereas the content of mito-
ferent genes. In contrast, alternative splicing of one chondria and the oxygen-carrying protein myoglobin

determines the endurance and overall color of the
muscle. White muscle cells depend largely on glycolysis
to supply ATP, accounting for their rapid fatigue com-
pared with red muscle cells, which are specialized for
oxidative metabolism with abundant mitochondria and
myoglobin.
Some muscles consist of only fast-twitch white muscle
cells or slow-twitch red muscle cells, but most muscles
are mixtures of two or more cell types. For example, in
chickens, the leg muscles that are responsible for sup-
porting the body, walking, and maintaining balance over
long periods of time are rich in red muscle cells (dark
meat). On the other hand, the chicken breast muscles,
used for energetic flapping of the wings for short periods,
are mainly white muscle cells (light meat).
Remarkably, the pattern of nerve stimulation deter-
mines the muscle cell type by controlling which genes
are expressed (and, presumably, how the troponin T
messenger RNA is processed). This was demonstrated
by transplanting motor nerves between fast and slow
muscles. Over a period of weeks, slow isoforms replaced
fast isoforms and vice versa. Even more surprising,
the same result is achieved by stimulating muscles
electrically with fast or slow patterns of impulses.
Chronic low-level stimulation biases gene expression
toward the proteins that are found in slow muscle cells.
Calcium and calmodulin provide one prominent link
between activity and gene expression. The concentration
of active calmodulin tracks with the pattern of stimula-
tion, because Ca2+ is released in the cytoplasm each
time a muscle contracts. Among other things, calcium-
calmodulin activates protein phosphatase PP2b (calcineu-
rin; see Fig. 25.6), which dephosphorylates transcription
factors (see Fig. 10.21). These activated transcription
factors move into the nucleus and help to establish a
transcription program that turns on expression of pro-
teins found in slow muscles. These include contractile
proteins and enzymes for oxidative metabolism.
The proportions of slow and fast muscle cells are
determined genetically, so world-class sprinters (with a
high proportion of fast, white fibers) and marathoners
(with a high proportion of slow, red fibers) are born with
advantages for their specialties. Training can lead to
hypertrophy of specific muscle cell types and improved
performance. Endurance training also leads to an
increased proportion of slow cells. Without training,
muscle strength declines with age; cell number remains
constant, but each cell decreases in size.
Structural Proteins of the Plasma Membrane:
Defects in Muscular Dystrophies
In addition to providing a permeability barrier, the
plasma membrane of the muscle cell must maintain its
integrity while being subjected to years of forceful
contractions. Occasional breaches of the membrane
are inevitable, so muscle cells also depend on a repair
CHAPTER 39 n Muscles 683
process that reseals holes. If membrane damage exceeds
the repair capacity, muscle cells degenerate locally (seg-
mental necrosis) or globally. Cell death beyond the
ability of muscle stem cells to regenerate the tissue
results in muscular dystrophy.
The proteins that stabilize muscle membranes were
discovered in the late 1980s, when mutations in the
dystrophin gene on the X-chromosome were linked
to Duchenne muscular dystrophy, the most common
human form of the disease. Dystrophin is an enor-
mous member of the -actinin superfamily of actin-
binding proteins (see Fig. 33.17). The dystroglycan
sarcoglycan complex (Fig. 39.17 and Table 39.3) was
found when it copurified with dystrophin after solubiliz-
ing the membrane with detergents.
More than 40 proteins are required to maintain the
integrity of the plasma membrane as shown by muta-
tions that cause muscular dystrophies (Table 39.3).
Disease-causing mutations in genes for proteins of the
dystroglycansarcoglycan complex typically lead to sec-
ondary loss of the other proteins in the complex. O-linked
glycosylation by Golgi apparatus glycosyltransferases is
A
Laminin
B
Sarcoglycan
complex Dystroglycan
Sarcospan complex
Plasma membrane
Dystrophin Syntrophin
complex
C Actin filament
FIGURE 39.17 DYSTROPHIN AND ASSOCIATED PROTEINS
STABILIZE THE PLASMA MEMBRANE OF SKELETAL MUSCLE.
AB, Fluorescent antibody staining of cross sections of human skeletal
muscle showing the localization of dystrophin at the plasma membrane
of a normal individual (A) and its absence in an individual with Duch-
enne muscular dystrophy (B). C, Model of the transmembrane complex
of proteins that links dystrophin and actin filaments in cytoplasm to
laminin in the basal lamina outside the cell. (AB, Courtesy L. Kunkel,
Harvard Medical School, Boston, MA. C, Based on a drawing by K.
Amann and J. Ervasti, University of Wisconsin, Madison.)
684 SECTION IX n Cytoskeleton and Cellular Motility

TABLE 39.3 Proteins Required to Stabilize and Repair Muscle Plasma Membranes
Protein Partners/Functions Expression Inheritance, Diseases
Membrane Skeleton
Dystrophin -Dystroglycan, actin Muscle, brain X-linked, DMD, BMD
Utrophin -Dystroglycan, actin Muscle, other tissues
-Syntrophins Dystrophin Muscle > other tissues None detected in humans
-Syntrophins Dystrophin, utrophin Muscle > other tissues None detected in humans
Transmembrane Proteins
Caveolin-3 Cholesterol Muscle AD, LGMD
-Dystroglycan Laminin, agrin Many tissues Embryonic lethal
-Dystroglycan Dystrophin, utrophin Many tissues Embryonic lethal
-Sarcoglycan Sarcoglycans, biglycan Muscle AR, LGMD, cardiomyopathy
-Sarcoglycan Sarcoglycans Muscle AR, LGMD
-Sarcoglycan Sarcoglycans, biglycan Muscle AR, LGMD
Integrin 7 Laminin Many tissues AR, CMD
Extracellular Matrix
Collagen VI 1, 2, 3 Biglycan Muscle, other tissues AD, Bethlem myopathy, Ulrich
syndrome
2-Laminin -Dystroglycan Muscle, other tissues AR, CMD, dy/dy mouse
Agrin -Dystroglycan, AChR Muscle Null mouse perinatal lethal
Sarcomeric Proteins
Titin Myosin, Z-disk Muscle AR, LGMD, tibial MD
Myotilin -actinin, Z-disk Muscle AR, LGMD
Golgi Enzymes That Process Membrane and ECM Proteins
Fukutin Glycosyltransferase Many tissues AR, Fukuyama CMD
LARGE Glycosyltransferase Many tissues AR, CMD
POMGnT1 Glycosyltransferase Many tissues AR, Muscle eye brain disease
POMTi O-mannosyltransferase Many tissues AR, Walker-Warburg syndrome
Membrane Repair Machinery
Dysferlin Muscle AR, LGMD, Miyoshi myopathy
Nuclear Envelope Proteins
Emerin Lamins, actin All cells XR, Emery-Dreifuss MD
Lamin A/C Nuclear envelope All cells AD/AR, LGMD, Emery-Dreifuss MD

AD, autosomal dominant; AR, autosomal recessive; BMD, Becker muscular dystrophy; CMD, childhood muscular dystrophy; DMD, Duchenne muscular
dystrophy; dy, dystrophia muscularis; ECM, extracellular matrix; LGMD, limb-girdle muscular dystrophy; MD, muscular dystrophy.

required for dystroglycan to bind to its extracellular
ligands including 2-laminin in the basal lamina. Proteins
in cytoplasmic vesicles are used to repair damaged
plasma membranes. The mechanical activity of muscle
cells might make them more sensitive than other cells to
deficiencies in proteins that support the nuclear enve-
lope (lamin A/C and emerin). Some of these mutations
also affect the nervous system.
Other than the X-linked dystrophin mutations, muta-
tions causing muscular dystrophies are usually autosomal
recessive. About one in several thousand humans devel-
ops some form of muscular dystrophy, because they
inherit mutations in both copies of one of the sensitive
genes. The mechanism of disease in muscular dystro-
phies is similar to that in hereditary spherocytosis, in
which deficiencies of the membrane skeleton make red
blood cells susceptible to mechanical damage (see Fig.
13.11). The age of onset and clinical features of inherited
muscular dystrophies depend on the molecular defect.
Patients with severe defects develop progressive muscle
weakness as children. Ultimately, failure of respiratory
muscles is fatal.
Dystroglycans and a dystrophin homolog, utrophin,
participate in clustering acetylcholine receptors at the
neuromuscular junction, the chemical synapse between
motor neurons and skeletal muscle (see Fig. 17.9).
When, during development, a motor neuron contacts
the surface of its target muscle cell, the neuron secretes
a proteoglycan called agrin, which is incorporated into
the adjacent basal lamina. Agrin binds dystroglycan and
a receptor tyrosine kinase in the muscle plasma mem-
brane, which position associated acetylcholine receptors
at the site where they receive acetylcholine secreted by
the nerve in response to an action potential.
 CHAPTER 39 n Muscles 685

Cell A

Cell B

Sarcomere Intercalated disk Gap junction

FIGURE 39.18 ELECTRON MICROGRAPHS OF A LONGITUDINAL SECTION OF TWO CARDIAC MUSCLE CELLS. Sarcomeres are
similar to skeletal muscle. Intercalated disks anchor neighboring cells together, and gap junctions couple the cells electrically. (Courtesy D.W.
Fawcett, Harvard Medical School, Boston, MA.)

Cardiac Muscle
To maintain the circulation of blood, heart muscle is Atrioventricular
specialized for repetitive (~100,000 times per day), node
Atria
fatigue-free contractions driven at regular intervals by
Sinoatrial
action potentials from specialized pace-making cells node
L
within the heart. Gap junctions allow these action poten-
tials to spread from one muscle cell to the next. Unlike R
skeletal muscle, the heart does not regenerate after
Ventricles
injury, so efforts are being made to reprogram cardiac
cells to recapitulate their normal development.

Contractile Apparatus of Cardiac Muscle Bundle

The architecture of the sarcomeres is very similar in
cardiac and skeletal muscle, but cardiac muscle has more
mitochondria, larger T tubules, and less SER (Fig. 39.18).
The human atrium has one major myosin isoform. Two
ventricular myosin isoforms (one shared with slow skel-
etal muscle) differ in ATPase activity and speed of con-
traction. Humans almost exclusively express only one of
these isoforms. Myosin-binding protein C binds at inter-
vals along the backbone of the thick filaments, interacts
with actin, and modulates the myosin cross-bridges. The
thin filaments are composed of a cardiac isoform of -
actin, tropomyosin, troponin, and a smaller version of
nebulin called nebulette. When the cells are damaged by
a heart attack or other disease, these proteins leak into
the blood. Cardiac isoforms TNI and TNT in blood are a
sensitive measure for cellular damage.
Short, modestly branched, cardiac muscle cells have
centrally located nuclei and squared-off ends (Fig. 39.1)
where cadherins (see Fig. 30.5) attach neighboring
cells to each other at specialized adhesive junctions
called intercalated disks (Fig. 39.18). These junctions
have features of both adherens junctions (links to
of His
Purkinje
fibers
FIGURE 39.19 ACTIVATION OF CARDIAC CONTRACTION. An
action potential (yellow arrows) starts at the sinoatrial node and travels
through atrial muscle cells to the atrioventricular node. After a short
delay at the atrioventricular node, the action potential spreads through
the interventricular septum in modified cardiac muscle cells, called
Purkinje fibers, and then through muscle cells to the whole ventricle.
The action potential follows the same path each time, giving rise to
electrical signals that can be detected on the body surface by electro-
cardiogram. Damage during myocardial infarctions changes the elec-
trocardiographic pattern and may cause arrhythmias.
actin filaments) and desmosomes (links to intermediate
filaments).
Pacemaker Cells
Intrinsically excitable pacemaker cells in the sino-
atrial node drive rhythmic contractions of the heart
(Fig. 39.19). The membrane potential of these cells drifts
spontaneously toward threshold, setting off action
potentials about once each second (Box 39.1). Each
686 SECTION IX n Cytoskeleton and Cellular Motility

BOX 39.1 Action Potentials in Cardiac Pacemaker Cells

Spontaneous action potentials of cardiac pacemaker cells in
the sinoatrial node are more complicated than those of
nerves (Fig. 39.20). Seven different plasma membrane chan-
nels determine their frequency:
1. Voltage-gated Na+ channels. As in nerves, these channels
rapidly activate at membrane potentials above threshold
and then rapidly inactivate.
1 m
2. T-type, voltage-gated Ca2+ channels. These low-
conductance channels activate transiently at membrane
potentials more negative than Na+ channels, about 70mV.
3. L-type, voltage-gated Ca2+ channels. These high-
conductance channels slowly activate and inactivate
when the membrane depolarizes to about 40mV. Sym-
pathetic nerve stimulation sensitizes these channels to
membrane depolarization. Drugs called dihydropyridines
block these channels.
4. Delayed-rectifier, voltage-gated K+ channels. As in nerves,
these HERG channels activate and inactivate slowly
in response to membrane depolarization. Sympathetic
0
nerves stimulate these channels.

Voltage (mV)
-15 5. Kir3.1 inward-rectifier K+ channels. These channels
conduct K+ over a limited range of membrane potential,
[Ca2+] i

-30

-45 between about 30 and 80mV. Parasympathetic nerve

-70
stimulation activates these channels.
2 3 2 3
0
7 1 4 7 1 4
6. Kir6.2 inward-rectifier K+ channels. Normal levels of
0 100 200 300 400
cytoplasmic ATP inhibit these channels. Depletion of
A Time (msec) cytoplasmic ATP activates these channels.
7. Nonselective K+/Na+ channels. Repolarization of the
membrane activates these HCN (hyperpolarization-
Na+ 1 Voltage-gated activated cyclic nucleotide-gated) channels, which slowly
Na+-channel
depolarize the membrane.
Acting together, these channels produce a spontaneous
2 T-type voltage-gated
Ca2+ cycle of pacemaker action potentials. At the threshold poten-
Ca2+-channel
tial (about 40 mV), voltage-gated Na+ channels open syn-
3 L-type voltage-gated chronously and rapidly depolarize the membrane. As they
Ca2+ Ca2+-channel inactivate, L-type Ca2+ channels open, prolonging the depo-
larization and admitting Ca2+; this, in turn, triggers contrac-
4 Voltage-gated tion by releasing more Ca2+ from internal stores (Fig. 39.14B).
K+ delayed-rectifier As these Ca2+ channels slowly inactivate, delayed-rectifier K+
K+-channel (HERG)
channels open and drive the membrane potential toward EK,
5 Kir3.1 inward-
the K+ equilibrium potential. As the membrane potential
K+
rectifier K+-channel reaches a minimum, delayed-rectifier K+ channels inactivate,
but the two Kir channels open. In the absence of other
K+ 6 Kir6.2 inward- channel activity, the membrane potential would remain near
rectifier K+-channel EK, but the nonselective Na+/K+ channels open and the mem-
brane slowly depolarizes, drifting toward threshold. T-type
7 Nonselective Ca2+ channels contribute to the slow, spontaneous depolar-
K+ Na+ K+/ Na+-channel
ization. At threshold, the cycle repeats.
B Channels similar to those in the sinoatrial node generates
action potentials in cardiac muscle cells and stimulate con-
FIGURE 39.20 MECHANISM OF SPONTANEOUS CARDIAC traction. Cardiac muscle cells can generate spontaneous
PACEMAKER ACTION POTENTIALS. Sequential activation and action potentials, but they have fewer T-type Ca2+ channels
inactivation of seven different plasma membrane channels account for
and more Kir K+ channels, so the rate of spontaneous action
the time course of action potentials and cytoplasmic Ca2+ transients
in pacemaker cells of the sinoatrial node. A, Time courses of the fluc-
potentials is lower than that of pacemaker cells. Except in
tuations in membrane potential (orange) and cytoplasmic Ca2+ con disease, pacemaker cells drive action potentials throughout
centration (blue). Colored boxes indicate when the various channels the rest of the heart. Later in life, cells in the pulmonary veins
enumerated in part B are open. Kir3.1 and Kir6.2 are inactive under can also generate action potentials. This is one cause of atrial
these conditions. B, Channels contributing to pacemaker activity. fibrillation, a common arrhythmia in older humans.

action potential spreads from cell to cell through gap
junctions (see Fig. 31.6), activating all cells in the atrium
within a few hundred milliseconds. After a brief delay in
the atrioventricular node, the action potential and
contraction spreads from cell to cell through the ven-
tricle. This highly reproducible pattern of electrical
activity can be recorded on the surface of the body as
an electrocardiogram.
Mutations in six different human ion channel genes
including voltage-gated sodium channels (SCN5A) and
potassium channels (HERG, KVLAT1, and minK) cause
disorders of cardiac muscle electrophysiology. These

A. Sympathetic stimulus
Norepinephrine
-Adrenergic Adenylyl
receptor cyclase
D
Gs T
Gs
Activates
Inhibits
Inactive cAMP Active
PKA
FIGURE 39.21 Regulation of the rate of cardiac pacemaker cells by sympathetic (A) and parasympathetic (B) nerves. GTP-Gs stimulates
adenylyl cyclase. GTP-Gi inhibits adenylyl cyclase. D is GDP associated with G protein -subunits; T is GTP. Ach, acetylcholine; cAMP, cyclic
adenosine monophosphate; GDP, guanosine diphosphate; GTP, guanosine triphosphate; PKA, protein kinase A.
inherited diseases are called long-QT syndrome because
the interval between the initial depolarization of the
muscle cells and their relaxation is prolonged. This
change predisposes the person to abnormal cardiac
rhythms that are potentially fatal.
Excitation Contraction Coupling
As in skeletal muscle, plasma membrane action
potentials stimulate cardiac muscle cells to contract
by releasing cytoplasmic Ca2+ to activate troponin-
tropomyosin (Fig. 39.14B). Calcium ATPase pumps (see
Fig. 14.7) in SER and plasma membrane maintain the low
cytoplasmic Ca2+ concentration with some help from
plasma membrane Na+/Ca2+ antiporters.
In both cardiac and skeletal muscle action potentials
activate L-type voltage-sensitive Ca2+ channels (DHP
receptors) in T tubules, but subsequent events differ as
revealed by a requirement for extracellular Ca2+ in heart
but not skeletal muscle (Fig. 39.14). Rather than interact-
ing directly with ryanodine receptors as in skeletal
muscle, the active cardiac L-type voltage-sensitive Ca2+
channels admit extracellular Ca2+. This opens nearby
ryanodine receptors in the SER, releasing a flood of Ca2+
to trigger contraction. This is called calcium-induced
calcium release.
Excitation-contraction coupling can be defective
when heart muscle cells grow larger in response to
abnormal demands, such as high blood pressure. The
defect may be explained by growth separating T tubules
from SER, either physically or functionally, thereby
decreasing the probability that Ca2+ entering through
DHP receptors will trigger Ca2+ release from the endo-
plasmic reticulum.
Seven-Helix Receptors and Trimeric G-Proteins
Regulate Heart Rate and Contractility
Motor nerves do not stimulate cardiac muscle directly.
Instead, molecules secreted by autonomic nerves
and the adrenal gland modulate the rate and force of
CHAPTER 39 n Muscles 687
B. Parasympathetic stimulus
Ca2+ K+ Acetylcholine
L-type Kir 3.1 Muscarinic Ach
Ca-channel K-channel receptor
G + GT GD
ATP
PKA
contraction (Fig. 39.21). Acetylcholine from parasym-
pathetic nerves slows the heartbeat, whereas norepi-
nephrine from sympathetic nerves and epinephrine
from the adrenal gland speed the rate and increase the
strength of contraction (Fig. 39.21). These neurotrans-
mitters target channels indirectly by activating two
different seven-helix receptors and their associated
trimeric G-proteins (see Fig. 25.9). The resting
rate reflects a compromise in the competition between
these two inputs.
Norepinephrine and epinephrine increase the heart
rate and force of contraction by modulating L-type Ca2+
channels (Fig. 39.21A). Both bind -adrenergic receptors
that activate trimeric G proteins, which stimulate the
production of cyclic adenosine monophosphate (cAMP)
by adenylyl cyclase (see Fig. 26.2). cAMP activates
protein kinase A (PKA) (see Fig. 25.3) that phosphory-
lates three types of channels. Phosphorylated L-type,
voltage-gated Ca2+ channels are more likely to open in
response to membrane depolarization and admit more
Ca2+ to activate the contractile machinery more fully.
Phosphorylated nonspecific HCN (hyperpolarization-
activated cyclic nucleotide-gated) cation channels push
the membrane potential toward threshold. Both increase
the frequency of action potentials and the heart rate.
Phosphorylated delayed-rectifier K+ channels are more
active in repolarizing the membrane, so they prevent
activated Ca2+ channels from prolonging the action
potential. Phosphorylation of phospholamban, the regu-
latory subunit of the calcium pump in the endoplasmic
reticulum, increases its activity, which speeds relaxation
These changes allows heart muscle cells to keep up with
stimuli generated at a higher rate from pacemaker cells.
PKA also targets sarcomeric proteins: phosphorylation
of troponin-I and myosin-binding protein C each increases
the rate of crossbridge cycling and the strength of
contraction.
Acetylcholine released by parasympathetic nerves
activates Kir3.1 inward-rectifier K+ channels that
688 SECTION IX n Cytoskeleton and Cellular Motility
slow the heartbeat (Fig. 39.21B). Acetylcholine binds
seven-helix receptors, called muscarinic acetylcholine
receptors (because they bind muscarine. These are dis-
tinct from pentameric nicotinic acetylcholine receptors
(see Fig. 16.12). Active muscarinic receptors catalyze the
exchange of guanosine diphosphate (GDP) for guano-
sine triphosphate (GTP) on the Gi subunit of a trimeric
G protein, releasing the G subunits to activate
Kir3.1/3.4 channels. When open, these channels reduce
the rate at which the membrane potential drifts toward
threshold. The free GTP-Gi subunits inhibit cAMP pro-
duction and reduce Ca2+ channel phosphorylation. This
decreases the probability that Ca2+ channels are open,
contributing to a lowering of the heart rate. If the energy
supply of the heart is compromised, ATP levels fall. This
activates Kir6.2 channels, which reduce the rate of spon-
taneous depolarization and lower the heart rate until
ATP levels are restored.
Therapeutic Effect of Digitalis in Congestive
Heart Failure
In congestive heart failure, cardiac contraction fails to
produce enough force to maintain adequate circulation
of blood. Digitalis from the foxglove plant was the first
drug to treat heart failure. Digitalis and related com-
pounds strengthen cardiac contraction indirectly by
inhibiting the 2 isoform of Na+K+-ATPase in the plasma
membrane (Fig. 39.22). Reduced sodium pump activity
lowers the Na+ gradient across the membrane, providing
less driving force for Na+/Ca2+ antiporters to exchange
extracellular Na+ for cytoplasmic Ca2+. The slightly
higher steady-state concentration of Ca2+ in cytoplasm
strengthens contraction.
3 Na+ 3 Na+ Ca2+
3 Na+ 3 Na+ Ca2+
Na+ /Ca2+
2 K+
Antiporter
ATP
ADP
Na+ K+ATPase
(target of cardiac
glycosides)
FIGURE 39.22 CHEMIOSMOTIC CYCLE THAT HELPS CLEAR
CA2+ FROM THE CYTOPLASM OF CARDIAC MUSCLE CELLS.
Na+K+-ATPase pumps create a Na+ gradient (purple triangle) to drive
the Na+/Ca2+ antiporter to transport Ca2+ up its concentration gradient
(blue triangle) out of the cell. Cardiac glycosides such as digitalis inhibit
the cardiac isoform of Na+K+-ATPase, raising the concentration of
cytoplasmic Ca2+ and strengthening cardiac contraction. K+ channels
(left) allow K+ to circulate.
Molecular Basis of Inherited Heart Diseases
Because the heart is so vital to survival, relatively minor
molecular defects command attention in humans. Nearly
1% of individuals carry an inherited or de novo mutation
in a gene for a sarcomeric protein that compromises
cardiac function. The most common mutations are in the
genes for myosin heavy chain, myosin-binding protein
C, and titin, but virtually every sarcomeric protein is
affected (Table 39.1). Long, noncoding RNAs participate
in regulating the stress response that leads to cardiomy-
opathies. Patients are typically heterozygous for these
mutations (ie, they are dominant negative mutations). In
hypertrophic cardiomyopathies, the heart attempts
to compensate for abnormal contractility (either
increased or decreased depending on the mutation)
through hypertrophy, but the thickened heart wall com-
promises cardiac relaxation and refilling the chambers
with blood. In dilated cardiomyopathies, the ventri-
cles swell and their walls thin. Both types are associated
with heart failure and abnormal cardiac rhythms that can
be fatal. The rate of progress of these diseases depends
on not only the particular mutation but also other factors
that vary from person to person. Individuals with defects
in myosin-binding protein C develop hypertrophy in
their fifties but can live normal life spans. By contrast,
those with defects in troponin T can be affected as teen-
agers and die of arrhythmias in their twenties. These
severe mutations of cardiac contractile proteins account
for about half of the deaths of apparently healthy young
athletes.
Smooth Muscle
Contractile Apparatus
Smooth muscle cells are specialized for slow, powerful,
efficient contractions under the control of a variety of
involuntary mechanisms. Smooth muscle cells are gener-
ally confined to internal organs, such as blood vessels
(where they regulate blood pressure), the gastrointestinal
tract (where they move food through the intestines), and
the respiratory system (where they control the diameters
of the air passages; their excessive contraction contrib-
utes to asthma and other allergic reactions). The cyto-
plasm of spindle-shaped smooth muscle cells (Fig. 39.1)
appears homogeneous by light microscopy, because the
contractile proteins are not organized in regular arrays
like sarcomeres of skeletal and cardiac muscle. A basal
lamina and variable amounts of collagen and elastic
fibers surround each cell. Smooth muscle cells rarely
divide in adults, but they are capable of responding
to local physiological conditions by remodeling their
structure, as in atherosclerosis and hypertension.
In terms of organization and biochemistry, smooth
muscle cells (Fig. 39.23) resemble nonmuscle cells more
than skeletal or cardiac muscle. For example, the gene
for smooth muscle myosin arose relatively recently from
a cytoplasmic myosin II gene. These myosins also share
 CHAPTER 39 n Muscles 689

Dense plaque
on plasma
membrane

Dense body
in cytoplasm

Myosin
filament

Intermediate
filament

C Actin filament

A
A B D
D

Dense
body
Myosin
Actin
IF

E
E

FIGURE 39.23 CONTRACTILE APPARATUS OF SMOOTH MUSCLE. A, Electron micrograph of a thin cross section of two smooth muscle
cells. BC, Organization of the contractile units, which stretch across the cell between plasma membrane attachment plaques. Contractile units
consist of myosin filaments connecting thin filaments attached to a dense body or plasma membrane plaque. D, High-power electron micrograph
showing a dense body and cross sections of three types of filaments. E, Electron micrograph of a longitudinal section of an extracted vascular
smooth muscle cell illustrating associations of actin filaments and intermediate filaments (IF) with dense bodies, and myosin filaments interacting
with actin filaments. (A, D, and E, Courtesy A.V. Somlyo and A.P. Somlyo, University of Virginia, Charlottesville. For reference, see Somlyo AP,
Devine CE, Somlyo AV, Rice RV. Filament organization in vertebrate smooth muscle. Philos Trans R Soc Lond B Biol Sci. 1973;265:223229;
Bond M, Somlyo AV. Dense bodies and actin polarity in vertebrate smooth muscle. J Cell Biol. 1982;95:403413.)

the same regulatory light chain. Long myosin thick fila-
ments are interspersed among the thin filaments, but not
in a regular way as in striated muscles. Thin filaments are
composed of actin and tropomyosin, along with two
regulatory proteins, caldesmon and calponin, rather than
troponin. Thin filaments are arranged obliquely in the
cell, some with their barbed ends attached to dense
plaques on the plasma membrane, others to dense
bodies in the cytoplasm. Like Z disks in striated muscles,
dense bodies anchor desmin intermediate filaments,
forming a continuous, inextensible, internal tendon
running from end to end of the cell, preventing excess
stretching (see Fig. 35.8).
Smooth muscle cells contract like a concertina (Fig.
39.24), because tension generated by myosin and actin
is applied to discrete spots on the plasma membrane.
This compression can be seen in light micrographs as
irregular cells with corkscrew nuclei. Given that
smooth muscle cells have less myosin than striated
muscle cells do, it is remarkable that they develop the
same force. This is explained by two factors. First, the
force-generating unit, the myosin filament, is larger in
690 SECTION IX n Cytoskeleton and Cellular Motility

Relaxed Contracted Regulation of Smooth Muscle Contraction

A wide range of stimuli trigger smooth muscle contrac-
tion, but they all seem to act through seven-helix recep-
tors coupled to trimeric G-proteins. Hormones stimulate
Stimulus
Force contraction of the uterus, whereas motor nerves stimu-
late intrinsic eye muscles that close the pupil.
Depending on the particular smooth muscle, Ca2+
Ca2+

Myosin-LC-P for contraction enters the cytoplasm through either

Ca2+ voltage-dependent calcium channels in the plasma mem-
brane or IP3 (inositol 1,4,5-triphosphate) receptor Ca2+
A Seconds release channels in the SER (Fig. 39.24). Drugs that block
plasma membrane calcium channels can distinguish
these two pathways experimentally. In intestines, para-
Stimulus MLCK sympathetic nerves release acetylcholine to stimulate
Calmodulin inactive seven-helix muscarinic receptors (Fig. 39.21). Associated
Ca2+ Ca-CM Ca2+ trimeric G-proteins activate cation channels that depolar-
ize the plasma membrane and allow Ca2+ to enter
MLCK through voltage-sensitive calcium channels. Conse-
active
IP3 quently, calcium channel blockers strongly inhibit activa-
Trimeric
G-protein tion of gut smooth muscle. Gap junctions couple gut
Myosin P Myosin
inactive active smooth muscle cells, allowing excitation to spread from
P
Phospholipase C cell to cell. At the other end of the spectrum, vascular
Myosin-LC smooth muscle depends on IP3 to release Ca2+ from intra-
Phosphatase
active cellular stores rather than depending on Ca2+ from
outside the cell.
RhoGTP Rho-kinase Phosphatase
Following stimulation, intracellular Ca2+ increases
P-MLC- rapidly but transiently, declining to a value above
Phosphatase
B inactive resting level as the receptors desensitize (see Fig. 24.3).
Ca2+ pumps in both SER and plasma membrane clear
FIGURE 39.24 ACTIVATION OF SMOOTH MUSCLE CON-
TRACTION. A, The spindle-shaped smooth muscle cell develops the cytoplasm of Ca2+ so that Ca2+ levels decrease to
accordion pleats as it contracts, owing to the attachment of the actin resting levels and the muscle eventually relaxes when
filaments at intervals along the plasma membrane. The graph shows the activating stimulus declines. Relaxing agents, acting
the time course of activation, consisting of the release of Ca2+ into the through cyclic guanosine monophosphate (cGMP) or
cytoplasm, phosphorylation of myosin regulatory light chains, and then
cAMP (see Fig. 26.1), promote clearance of cytoplas-
the slow development of force. Myosin light-chain phosphorylation
(LC-P) is required to initiate, but not to sustain, the contraction of mic Ca2+. Epinephrine relaxes smooth muscles of the
smooth muscle. B, Biochemical pathways controlling phosphorylation respiratory system by another mechanism. Stimulation
of myosin regulatory light chains. Receptor stimulation leads to pro- of -adrenergic receptors activates potassium channels
duction of IP3 (inositol 1,4,5-triphosphate) by phospholipase C and that hyperpolarize the plasma membrane and reduce
release of Ca2+ into cytoplasm. Ca2+ binds calmodulin (CM), which
Ca2+ entry. This approach is widely used to treat asthma.
activates myosin light-chain kinase (MLCK) by binding the kinases
autoinhibitory peptide and displacing it from the active site. Active After a considerable delay (>200ms) following
MLCK phosphorylates activating sites on the regulatory light chain. the Ca2+ spike, contractile force develops slowly. The
Light-chain phosphatase reverses phosphorylation of myosin. Activa- delay is attributable to the time required for a sequence
tion of the small GTPase (guanosine triphosphatase) Rho with GTP of three biochemical reactions: Ca2+ binding to calmodu-
stimulates Rho-kinase, which phosphorylates and inactivates light-
lin, calcium-calmodulin activation of myosin light-
chain phosphatase. This makes the system more sensitive to Ca2+
levels, as light-chain phosphorylation is prolonged. LC, light chain; chain kinase (see Fig. 25.4), and phosphorylation of
P-MLC-, phosphorylated myosin light chain. (A, Modified from the myosin regulatory light chains, turning on the myosin-
work of K. Kamm and J. Stull, University of Texas Southwestern actin ATPase cycle (Fig. 39.24). Unphosphorylated
Medical School, Dallas.) myosin-II from smooth muscle and vertebrate nonmus-
cle cells is inactive.
smooth muscle than in skeletal muscle. Deploying a Phosphorylation of myosin light chains is required to
given amount of myosin in large, thick filaments in a long initiate but not maintain contraction, so slowly cycling,
sarcomere produces more force than does the same unphosphorylated myosins maintain peak force with
myosin in smaller filaments arranged in a series of short little expenditure of energy. Regulation of unphosphory-
sarcomeres. Second, individual smooth muscle myosin lated crossbridges is not well understood, but they
molecules produce a larger force than skeletal muscle appear to be activated cooperatively by a small popula-
myosin, at least in vitro assays. tion of phosphorylated myosin heads. Caldesmon, a

calcium-calmodulin-binding protein associated with
tropomyosin on actin filaments, may contribute to activa-
tion and/or allow myosin heads to cycle very slowly even
in the presence of ATP.
The sensitivity of light-chain phosphorylation to Ca2+
depends on a parallel signaling pathway that partially
inhibits myosin phosphatase, thus increasing the number
of phosphorylated myosin cross-bridges and force at
any given Ca2+ concentration (Fig. 39.24). Receptors
coupled to trimeric G-proteins activate the small
guanosine triphosphatase (GTPase) RhoA, which
stimulates a protein kinase that inhibits myosin light-
chain phosphatase.
ACKNOWLEDGMENTS
We thank Lee Sweeney and John Solaro for their sugges-
tions on revisions to this chapter.
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