PHYSIOLOGY SGD 3: SIGNAL TRANSDUCTION

1. Discuss figure 3-1 of Berne and Levi 6

th
ed. and relate it to the meaning of signal transduction.

Signal transduction common way wherein cells communicate with each other via chemical messengers and ligands
The sequence of events between receptor activation by a messenger and cellular response


2. Differentiate the types of plasma membrane receptors based on the signal transduction pathway used.

Plasma Membrane Receptors (water soluble)
A. Receptors that are ligand gated ion channels
PATHWAY:
When ligand binds to the channel receptors, the pore of the channel opens, allowing ions through the channel
When ligand dissociates, the pore once again closes
Usually NT activated
e.g. 1. Nicotinic Ach receptor
The ligand is Ach, Na ions are gated by AchR
The entry of Na will cause a change in membrane potential -> AP
2. Opening of ligand gated Ca channels cause by the entry of Ca
Ca acting as a second messenger binds to calmodulin which will then activate the downstream proteins leading to
a cellular response

B. Receptors that function as enzymes (e.g. receptor tyrosine kinase)
Possesses intrinsic enzyme activity (protein kinases)

Classified based on MOA:
o Receptor Tyrosine Kinase
PATHWAY:
Ligand binding causes activation (conformational change) of the receptors intracytoplasmic/enzymatic
portion
Receptor phosphorylates portion of proteins that contain tyrosine groups
Phosphotyrosine formed serves as a docking site for cytoplasmic proteins
Bound docking proteins bind and active other proteins which in turn activate more signaling pathways
Cascade of protein activation ending in a cellular response
e.g. 1. EGF binds with receptor tyrosine kinase to stimulate proliferation and differentiation of epithelial cells
2. Mutations in fibroblast growth factor receptors -> congenital skeletal abnormalities (dwarfism)

o Receptor Guanylyl Cyclase
PATHWAY:
When stimulated, receptor guanylyl cyclase is activated
The receptor enzyme complex cleaves or dephosphorylates GTP to form cGMP
cGMP acts as a second messenger which activates protein kinase G (PKG)
PKG activates the downstream effector proteins leading to a cellular response
e.g. ANP binds to receptor guanylyl cyclase -> excretion of Na and water by the collecting ducts of the kidneys
*also causes relaxation of the smooth muscle in the heart

C. Receptors that are bound to and activate cytoplasmic janus kinases
Enzymatic activity does not reside in the receptor, but in separate cytoplasmic kinases janus kinases (JAKs)
Receptor and its associated JAK functions as a unit
PATHWAY:
Ligand binding causes conformational change in the receptor
The receptor activates the attached JAK kinase
JAK phosphorylate target protein to act as transcription factors that will lead to synthesis of new proteins
JAK activates the downstram proteins triggering a cascade of protein activation leading to a cellular response
e.g. Erythropoietin binds with cytokine receptors with JAK kinase to promote growth and differentiation of red cell precursor

D. G-protein coupled receptors that activate G-proteins (GPCR)
Classified accdg to the type of secondary messengers it produces
cAMP
cGMP
IP3 & DAG
Arachidonic acid

COMPONENTS OF SIGNAL TRANSDUCTION PATHWAY INITIATED BY GPCR STIMULATION:
Ligand
Receptor
G-protein
Effector protein
Second messenger
Protein kinase
Downstream proteins

G-protein is attached to the receptor (trimeric GTP binding protein)
o Alpha binds and hydrolyzes GTP
o Beta forms a stable, tight non-covalent link
o Gamma
Exists in two forms based on what is attached: GDP/GTP
INACTIVATED/Resting G-protein
Has GDP attached to the alpha subunit
Remains as a single protein
ACTIVATED G-protein
Attached GDP is replaced by GTP
G-protein dissociates into 2 components
o Beta-gamma component
o Alpha component

3. Tabulate the signal transduction pathways utilized by the GPCR via cAMP, cGMP, and DAG second messengers.

Ligand binding
G-protein attaches to the receptor
GTP replaces GDP
G-protein detaches from the receptor and subunits disassembles
Subunits attach to the effector protein
GDP replace GTP
G-protein reassembles

cAMP
Ligand (1
st
messenger) binds to its specific receptor
Receptor activation leads to G protein (s/i type) dissociation
Alpha subunit activates the adenyl cyclase (1
st
effector protein)
Adenyl cyclase converts ATP to cAMP (2
nd
messenger)
PKA either
o Stimulate other downstream proteins -> cellular response
o Goes into the nucleus to stimulate DNA transcription -> mRNA formation & translation -> cellular response
cAMP stimulation of the downstream event is terminated by:
o ligand dissociates with the receptor
GDP replacing GTP
Rebinding of alpha subunit with the beta gamma subunit -> cessation of adenyl cyclase activity
o cAMP is broken down by enzyme phospodiesterase
Beta gamma subunit is not totally inert. In some cells it participates in the regulation of signal transduction
e.g. epinephrine effect on liver cells to release glucose

cGMP
Ligand (1
st
messenger) binds to its specific receptor
Receptor activation leads to G protein (t type) dissociation
Alpha subunit activates cGMP phosphodiesterase (1
st
effector protein)
Phosphodiesterase breaks down cGMP (2
nd
messenger) to GMP
Decrease cGMP leads toclosure of cGMP dependent cation channels
Ensuing changes in cation channel activity alters membrane voltage
e.g. light rays acting on photoreceptor protein of the eye (transducin)

IP3/DAG
Ligand (1
st
messenger) binds to its specific receptor
Receptor activation leads to G-protein (q type) dissociation
Alpha subunit activates phospholipase C (1
st
effector protein)
Phospholipase C breaks down PIP2 (phosphatidylinositol diphosphate) to produce:
o IP3 (Inositol triphosphate)
o DAG (Diacylglycerol)
Steps after IP3:
IP3 stimulates a ligand gated calcium channel on the endoplasmic reticulum into the cytoplasm to open up
Ca moves out of the ER into the cytoplasm
Ca binds to calmodulin
Ca-calmodulin stimulates protein kinase C
Protein kinase C stimulates downstream proteins -> cellular response
Steps after DAG:
DAG stimulates protein kinase C
Protein kinase C stimulates downstream proteins -> cellular response
e.g. TSH, Vasopressin, GABA

Arachidonic Acid
Ligand (1
st
messenger) binds to its specific receptor
Receptor activation leads to G-protein dissociation
Alpha subunit activates phospholipase A2 (1
st
effector protein)
Phospholipase A2 breaks down phospholipid to form arachidonic acid
Depending on the enzymes present in the cell, the AA is further converted into 3 different pathways to produce several types of
2
nd
messengers (Eicosanoids):
Cyclooxygenase Thromboxanes, Prostacyclins, Prostaglandins
Lipooxygenase 5 HETEs, Leukotrienes
Epoxygenase HETEs, EETs

cAMP cGMP IP3/DAG AA
G-protein Gs/Gi Gt Gq
Effector protein Adenyl cyclase PDE Phospholipase C Phospholipase A2
Second messenger cAMP cGMP IP3/DAG AA, Eicosanoids
Protein kinase PKA PKG PKC COX thromboxanes,
prostaglandins,
prostacyclins
Lipooxygenase 5HETE,
leukotrienes
Epoxygenase HETEs,
EETs

4. Explain how extremely minute amount of hormone epinephrine can be effective in alleviating symptoms related to hypoglycemia.
*Epinephrine effect on liver cells -> to release glucose
Epinephrine is synthesized by adrenal gland

5. In the disease cholera, its toxin enters the intestinal cells to modify G proteins.
A. What will be the expected effect on the G protein? Explain if it will be activated or not.
The G-protein will be chemically modified by the toxin? It will be inactivated.


B. What membrane signal transduction process is improperly regulated in cholera? What will be the effect on the signal transduction
pathway?
Plasma membrane G-proteins? G-proteins can no longer cleave GTP into GDP = no cellular response?
Effect on signal transduction pathway blocked/stopped?

C. What biological effect is expected?
Severe diarrhea?


6. An asthmatic patient was prescribed with inhaled steroids and salbutamol. He took the steroids as prescribed but he took the
salbutamol more than the frequency prescribed by the doctor. Initially the salbutamol afforded the relief of shortness of breath
however weeks later he noticed it was not as effective during the initial week of intake.

A. Enumerate the different components of the specific signal transduction pathway utilized by salbutamol to relax bronchial smooth
muscles.

*? Directly acting adrenergic agonists act on adrenergic receptors. All adrenergic receptors are G-protein coupled, activating signal
transduction pathways. The G-protein receptor can affect the function of adenylate cyclase or phospholipase C, and agonist of the
receptor will up-regulate the effects on the downstream pathway (it will not necessarily up-regulate the pathway itself).

Binds to the B2 receptor, induces a conformational change in G-proteins -> muscle relaxation

B. Using the concepts of up regulation and down regulation receptors, explain why salbutamol was ineffective during the later part
of treatment.

Up-regulation adaptation of the cell wherein the total number of plasma membrane receptors is increased due to a chronic low
concentration of the ligand
Down regulation adaptation of the cell wherein the total number of plasma membrane receptors is diminished due to a chronic
high concentration of its ligand

DOWN-REGULATION
The smooth muscle has been exposed to a high concentration of B2 agonist Salbutamol which leads to 100% receptor
saturation and maximal smooth muscle relaxation.
The smooth muscle may perceive this as detrimental to its function and therefore initiates a process known as B2 receptor
complex endocytosis.
This will decrease the number of receptor stimulated consequently leading to lesser second messenger formation and
lesser smooth muscle relaxation.