TOPICS FOR TODAY

 Gt transducing
 GPCR ADAPTATION
 Enzyme Linked Receptor
 Receptor Tyrosine kinase
 IP₃-DAG Pathway
 RAS-MAP KINASE PATHWAY
 JAK/STAT
 SER/THR KINASE PATHWAY
 NITRIC OXIDE
Gt Transducing
• Vision depends on G-protein coupled receptors that regulate cyclic nucleotide gated channels.

Rhodopsin is a receptor 7 membrane 11-cis Retinal light absorbing

spanning α-helices present in rod cells. chromophore Opsin-the
protein
component

11 cis Retinal 11-trans Retinal

• TRANSDUCIN-Gt
DARK-GDP-NO SIGNAL-cyclic GMP
Effector Molecule- cyclic GMP Phosphodiesterase
1. Light absorption converts 11-cisretinal to all-transretinal, activating
rhodopsin (Rh).
2. Activated rhodopsin catalyzes replacement of GDP by GTP on transducin
(T), which then dissociates into Tα-GTP and Tβγ
3. Tα-GTP activates cGMP phosphodiesterase (PDE) by binding and removing
its inhibitory subunit (I).
4. Active PDE reduces [cGMP] to below the level needed to keep cation
channels open.
5. Cation channels close, preventing influx of Na+ and Ca2+; membrane is
hyperpolarized. This signal passes to the brain.
6. Continued efflux of Ca2+ through the Na+-Ca2+ exchanger reduces
cytosolic [Ca2+]
7. Reduction of [Ca2+] activates guanylyl cyclase (GC) and inhibits PDE;
[cGMP] rises toward “dark” level, reopening cation channels and returning
Vm to prestimulus level.
8. Rhodopsin kinase (RK) phosphorylates “bleached” rhodopsin; low [Ca2+] and
recoverin (Recov) stimulate this reaction. Arrestin (Arr) binds
phosphorylated carboxyl terminus, inactivating rhodopsin.
9. Slowly, arrestin dissociates, rhodopsin is dephosphorylated, and all-
transretinal is replaced with 11-cis-retinal. Rhodopsin is ready for another
phototransduction cycle.
 Conformational
change of Rhodopsin

Decreased Release of Activation of

synaptic transmitter transducin

Activation of
hyperphosphorylation
phosphodiesterase

Decreased
Closure of Na+
intracellular cyclic
Channels
GMP
GPCR ADAPTATION
• Diminution of receptor signalling in the presence of continued or repeated
stimulation.

desensitization Internalisation

• G-Protein COUPLED RECEPTOR KINASE(GRKs) Arrestin Uncouple receptor

and G-Protein exocytosis
• In internalisation- removal of receptors from cell surface degrade
• RECEPTOR SEQUESTRATION-intracellular membranous compartments
• RECEPTOR DOWNREGULATION- degrade in lysosomes(hours)
 Classes
G-Protein
Coupled Ion-Channel
Receptors (GPCR) Receptors

Enzyme linked
receptors
• Receptor TYROSINE KINASE

• TYROSINE KINASE ASSOCIATED

RECEPTORS

• RECEPTOR SERINE/THREONINE
Receptor tyrosine Kinase
These receptors have an intrinsic enzyme activity instead of G-protein.

LIGANDS-
Insulin, Epidermal
Growth factor(EGF),
Transforming growth
factor (TGFα),Platelet
Derived Growth
Factor(PDGF),fibroblast
growth factor(FGF),
Colony Stimulating
Factor (CSF)
RAS-MAP Kinase Pathway
• RAS is an intracellular monomeric GTPase switch protein.
• It is a lipid linked protein present at the cytoplasmic face of the membrane.

GTPase ACTIVATING
PROTEIN (GAP)

GTP GDP

GUANINE NUCLEOTIDE
EXCHANGE FACTOR (GEF)

GRB2-An Adapter Protein

GEF- SOS
MAP-KINASE PATHWAY

• MAP-Mitogen Activated Protein Kinase

• Regulates cell-cycle and differentiated –specific proteins
• Serine/threonine kinase phosphorylation cascade

RAF MEK
ERK
MAP-kinase-kinase - MAP-kinase-
MAP- kinase
kinase kinase

• Effect is change in the pattern of transcription

• RHO-Present in cytosol
IP₃-DAG Pathway
• Activates phospholipase-Cγ. It binds the SH2 Domains binds to
phosphotyrosine.

PHOSPHATIDYLINOSITOL Inositol 1,4,5-

4,5-bisphophate (PIP₂) triphosphate (IP₃)

diacylglycerol
PI-3 Kinase Pathways
• PKB a Protein Kinase B –ser/thr kinase and PH domain binding
3phosphate of PI-3,4,5-triphosphate
• PKB(Akt) present in cytosol-inactivate binds PIP3- ACTIVATE
Insulin receptor contains two α (N terminal) and β (C-terminal)
subunits.
Glycogen synthase kinase Phosphorylated- Inactive
Glycogen Synthase Phosphorylated- Inactive
Tyrosine Kinases Associated Receptor
• Stimulate intracellular enzymes with which they non-covalently
associated. C-terminal is devoid of any known catalytic activity so
associate with CYTOSOLIC tyrosine kinase.
Most cytokines-IL-2 and erythropoietin.
Polypeptide hormone-growth hormone and prolactin Receptors mainly for
antigens, cytokines,
interleukins,
polypeptide
hormone
 Serine/Threonine Kinase Receptor

• Receptors- Type I Type II

• Homodimer of each type I and type II forms heterodimer

• Type I is activated by type II which then activates smad
• Smad transfer into nucleus and function to control gene expression.
Ligands- TGF-β
superfamily
including ACTIVINS,
BONE
MORPHOGENETIC
FACTOR (BMP)
Nitric Oxide
• It is a PARACRINE SIGNALING MOLECULE. Gaseous ligand affecting
nearby cell and highly unstable.
• It can diffuse the plasma membrane.
• It relaxes smooth muscle-dilation of blood vessels
• Here PKG plays a role
• EFFECTOR ENZYME- Guanyl cyclase
Questions
• GPCR consists of three subunits α, β, γ. I n unstimulated state α is
bound to GDP and GPCR is inactive. When GPCR is activated it acts
as GEF and allows α subunit to bind GTP and remove GDP. In order
to regulate GTP/GDP concentration α-subunit acts as
1. GTPase
2. GDP kinase
3. cGMP phosphodiesterase
4. cAMP phosphodiesterase