5.

Molecular motors

Prof. G. Gilardi - Biological Chemistry

1
 MOTILITY CAN CONCERN THE ENTIRE
ORGANISM, PART OF IT, SINGLE CELLS
OR SUBCELLULAR COMPONENTS.

ELEMENTS FOR MOTILITY.

MUSCLES: ACTIN-MIOSIN SYSTEM

SUBCELLULAR COMPONENTS
(CHROMOSOMES, CILIA AND FLAGELLA):
PROTEINS-MICROTUBULES
INTEARACTION = TUBULIN

MOLECULAR MOTORS: RNA

POLYMERASE.
Prof. G. Gilardi - Biological Chemistry 2
 All the biological systems involved in
movement have common features:
1. ATP hydrolysis is converted into mechanical work.
2. Movements are generated from motions in protein
molecules: energy transducers = chemical energy
mechanical work.
There are 3 main types of molecular motors:
1. Actin-myosin contractile system
2. Microtubule systems: tubulin, dynein, kinesin
3. Bacterial flagella: flagellin
Prof. G. Gilardi - Biological Chemistry 3
 ACTIN
Under physiological conditions, actin exists as:
1. A long, helical polymer = fibrous actin (F-actin)
2. A globular protein monomer (G-actin).
The structure of the G-actin monomer is a two-domain molecule with a mass of 42,000 Daltons.
The binding of ATP by a G-actin
monomer leads to polymerization, that
Is the formation of F-actin
In F-actin, the G-actin monomers are arranged in a 2-stranded helix
The 2 ends of the F-actin strands are asymmetric: there is a plus end
and a minus end
The actin filaments carries sites on each monumer that can bind myosin

G-actin F-actin

Prof. G. Gilardi - Biological Chemistry 4

Prof. G. Gilardi - Biological Chemistry 5
 MYOSIN
The functional myosin molecule is composed of six polypeptide
chains: two identical heavy chains (M = 230,000) and two each of two
kinds of light chains (M = 20,000)

The heavy chains have long -helical tails

and globular head domains.
The -helical tails are interwound into a
two-strand coiled coil and the light chains
are bound to the globular head domains.
Between each head domain and tail
domain is a flexible stalk.

Prof. G. Gilardi - Biological Chemistry 6

 CLEAVAGE BY PROTEASES

The tail domain can be cleaved at a specific point

by trypsin to yield fragments called light meromyosin

and heavy meromyosin.

Myosin exhibits aspects of both fibrous and globular

proteins, and its functional domains play quite different
roles.

The tail domains have a pronounced tendency to

aggregate, causing myosin molecules to form thick
bipolar filaments.

The head domains, with their attached light chains, are

often called headpieces; they have a strong tendency to
bind to actin.
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Myosin: assembly and
S1 structure

Prof. G. Gilardi - Biological Chemistry 8

 ACTIN-MIOSIN
INTERACTION
Actin filament is
surrounded by S1 heads.

The filament has a

direction because of the
asymmetry.

In presence of actin, the

S1 fragment have ATP-
ase activity.

The hydrolysis of ATP

breaks the actin-miosin
interaction.

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Prof. G. Gilardi - Biological Chemistry 10
4. Release of phosphate and ADP from a previously
hydrolyzed ATP on the thick filament causes it to
perform a "power stroke", pulling the thin filament
(actin) in towards the center of the sarcomere.
5. The binding of ATP causes the cross-bridge
between actin and myosin to be broken and the
myosin headpiece to remain in the low energy
configuration. In the absence of calcium, the actin
can slide back past the myosin to the original relaxed
position.
6. ATP is hydrolyzed, but not released by the myosin
headpiece, causing the myosin headpiece to
assume the cocked position.
7. When the stimulation from the nervous system
ceases, calcium is taken up again by the
sarcoplasmic reticulum.
8. The resulting decrease in calcium concentration
causes the thin filament to slide back past the thick
filament an

Prof. G. Gilardi - Biological Chemistry 11

 Energetics
Force of 1 power stroke 5 pN = 10-20 J = 1/5 energy release from the
hydrolysis of 1 ATP molecule
1 power stroke = 10-20 nm
1 ATP = 100 nm

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 Energy storage:
ATP is an intermediary energy storage,
Creatine phosphate is the long term storage
molecule

Prof. G. Gilardi - Biological Chemistry 13

 The Sliding
Filament Model
The sliding filament model explains the
molecular basis by which muscular
contraction occurs. During muscular
contraction, thin filaments within the
sarcomere of a myofibril are pulled towards
the center of the sarcomere (called the H
zone) by the thick filaments. In the process,
the sarcomeric length shortens and the
myofibril shortens. As a result, the muscle
contracts
Prof. G. Gilardi - Biological Chemistry 14
 The role of Ca 2

1. Muscular contraction is initiated by a signal from the

nervous system.
2. The nervous system causes release of calcium from
the sarcoplasmic reticulum.
3. Calcium release causes thick filaments of the cocked
headpiece of a myosin filament (thick filament) to
bind to a site on actin, forming a tight cross-bridged
binding. In the absence of calcium from the
sarcoplasmic reticulum, access of the headpiece to
the thin filament is blocked by tropomyosin and the
troponins (I, C, and T). Calcium binding by troponin
C causes a rearrangement of the troponin-
tropomyosin-actin complex, allowing actin-myosin
cross-bridges to form.

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Ca 10-7 M Ca 10-5 M

Prof. G. Gilardi - Biological Chemistry 16

Thin filaments are composed of a polymer of
actin (called F-actin) arranged in a helix,
tropomyosin (a fibrous protein that exists as elongated
dimers lying along, or close to, the groove in the F-actin
helix),
three small proteins called troponins I, C, and T.
The presence of tropomyosin and the troponins inhibits the
binding of myosin heads to actin unless calcium is present
at a concentration of about 10-5 M. In resting muscle,
calcium concentrations are approximately 10-7M, so new
cross bridges between the thick and thin filaments cannot
occur. Prof. G. Gilardi - Biological Chemistry 17
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Non-muscle actin
and myosin

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 Microtubule Systems

Microtubules are systems of

very long tubular structures built
from a helical wrapping of tubulin
Tubulin is involved in motility
within cells, such as in the mitotic
spindle, flagella, and nerve
axons.
Tubulin is composed of two
subunits, and , each of
molecular weight 55,000. They
are present in equimolar
amounts in tubulin as dimers.

Prof. G. Gilardi - Biological Chemistry 21

 Assembly and MAPs
Assembly of microtubules requires
energy from the hydrolysis of GTP.
One end of the microtubule grows
more rapidly, and is called the plus
end.
The final assembly of a functional
microtubule usually involves the
binding of other proteins, called
microtubule-associated proteins
(MAPs) to its surface.
One important MAP is the tau-MAP:
Phosphorylation of tau-MAP causes
dissociation from microtubules with their
destabilisation
Hyper-phosphorylation forms tangled of
tau-filaments in neuronal axons leading to
Alzheimers disease

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Cilia and flagella

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 Bacterial Motility
Rotating Proteins
The bacterial flagellum is a right-hand
helical fiber, composed almost
entirely of the fibrous protein flagellin.
That is, the flagellum does not contain
microtubules, actin, myosin
The flagellum itself rotates as a
means of propelling the bacterium
The driving force for rotation of the
flagellum is a gradient of protons
moving across the bacterial inner
membrane. Thus, the flagellum
behaves like an electric motor, in a
sense. The motor runs at about 100
revolutions per second and requires
the passage of about 1000 protons
per revolution. Marine bacteria have
similar rotary motors, but they use a
sodium gradient, not a proton
gradient, to power the motor.
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Prof. G. Gilardi - Biological Chemistry 28
Chemotaxis

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