G-protein linked receptors

Diversity of GPCRs

The Major Families of Trimeric G proteins*

---------------------------------------------------------------------------------------------------------------Family Members Action mediated by Functions ---------------------------------------------------------------------------------------------------------------I Gs activates adenylyl cyclase; activates Ca2+ channels Golf activates adenylyl cyclase in olfactory sensory neurons II Gi inhibits adenylyl cyclase activates K+ channels Go activates K+ channels; inactivates Ca2+ channels and activates phospholipase C- Gt (transducin) activates cyclic GMP phosphodiesterase in vertebrate rod photoreceptors III Gq activates phospholipase C- ---------------------------------------------------------------------------------------------------------------IV G12 G13 activates chloride channel activates Na+/H+ exchanger pathways activates small G protein Rho family, leading to actin cytoskeletal reorganization

G-protein-linked receptors; functionally diverse but share a common structure

extra-cellular
Ligand binding

G-protein-linked receptors function through trimeric G-proteins

G-protein-linked receptors
mediate their intracellular actions through target ion channels or enzymes. pathway always involves activation of one or more guanine nucleotide-binding regulatory proteins (trimeric G proteins).
trimeric G proteins consist of three protein subunits; alpha, beta and gamma. The G alpha binds a guanyl nucleotide. Various types, each specific for a set of serpentine receptors and for a particular downstream target, but they all operate the same way.

Receptor, inactive G-proteins, and adenylyl cyclase are within shouting distance in the cell membrane.

Ligand binding (1) -conformational change in the receptor -separates TMs. -separation of the TMs may open a crevice for binding to G.
Receptor binds to G protein (2)

Receptor activation results in activation of adenylyl cyclase. -indirect -stimulates a trimeric G-protein -trimeric G-proteins dissasemble when activated. Receptor binds to G-protein induces conformational change (3) GDP is replaced by GTP G dissociates from G The binding site for adenylyl cyclase is unmasked. G then binds to adenylyl cyclase (4), activating synthesis of cAMP

A single hormone/ receptor complex stimulates the production of many molecules of Gs

The binding of the Gs subunit to adenylyl cyclase activates the enzyme to produce many molecules of cAMP.

signal amplification
Binding of G to adenylyl cyclase causes a conformation change in G and GTP is hydrolyzed to GDP. This causes G to dissociate from adenylyl cyclase and re-bind G

Terminating the response

The hormone/receptor complex must be deactivated to return to the unstimulated state. -phosphorylation events on the carboxy terminal tail of the receptor lead to the inactivation of the receptor. Hydrolysis of GTP leads to inactivation of the trimeric G-protein. -enhanced by RGS proteins (regulator of G-protein signaling).

The trimeric GTP-binding proteins act as molecular switches

Functionally couple the G-protein-linked receptors to their target enzymes or ion channels. GTPases that function as molecular switches -flip between two states: active and inactive. -Inactive; trimer bound to GDP through G -Active; G bound to GTP

two conformation states

active

A G-protein which acts to stimulate a target enzyme is called a G stimulatory (Gs). Gi is inhibitory.

inactive

Mammalian RGS proteins activate the GTPase activities of G-protein alpha subunits
RGS proteins are GAPs (GTPase activating proteins). -no effect on the time course of nucleotide binding -but they stimulate the rate of GTP hydrolysis.
GTP GTP hydrolysis

GDP

RGS protein

MODEL: RGS proteins accelerate GTP hydrolysis by preferentially binding to and stabilizing G proteins in their transition state for the hydrolysis reaction.

Inhibitory G proteins
While the beta-adrenergic receptors are functionally coupled to G-stimulatory proteins, the alpha-2 adrenergic receptors are coupled to inhibitory G proteins. Gi can contain the same beta/gamma subunits as Gs, but the alpha subunits are different. Gi inhibits adenylyl cyclase in an indirect manner.

Hormone-induced activation and inhibition of adenylyl cyclase is mediated by G-s and G-i

GPCR-mediated Signaling Pathways

out membrane Gs AC PLC-

PIP 2

in

Ras/ Raf PI3K ATP cAMP Rho MEK AKT

MAPK

AC ATP cAMP

DAG+IP 3

Rac

[Ca+2]i PKC activation mobilization

proliferation survival

actin cytoskeletal rearrangement cell cycle progression

GPCR Classes
Class A: Rhodopsin like Class B: Secretin like Class C: Metabotropic glutamate / pheromone Class D: Fungal pheromone Class E: cAMP receptors Frizzled/Smoothened family Putative families: * Ocular albinism proteins * Insect odorant receptors * Plant Mlo receptors * Nematode chemoreceptors * Vomeronasal receptors (V1R & V3R) * Taste receptors T2R Orphans: * Putative / unclassified GPCRs non-GPCR families: * Class Z: Archaeal/bacterial/fungal opsins

GPCR Classes
* Class A Rhodopsin like o Amine o Peptide o Hormone protein o (Rhod)opsin o Olfactory o Prostanoid o Nucleotide-like
o Cannabinoid o Platelet activating factor o Gonadotropin-releasing hormone o Thyrotropin-releasing hormone & Secretagogue o Melatonin o Viral o Lysosphingolipid & LPA (EDG) o Leukotriene B4 receptor o Class A Orphan/other * Class C Metabotropic glutamate / pheromone o Metabotropic glutamate o Calcium-sensing like o Putative pheromone receptors o GABA-B o Orphan GPRC5 o Orphan GPCR6 o Bride of sevenless proteins (BOSS) o Taste receptors (T1R) * Class D Fungal pheromone o Fungal pheromone A-Factor like (STE2,STE3) o Fungal pheromone B like (BAR,BBR,RCB,PRA) o Fungal pheromone M- and P-Factor * Class E cAMP receptors * Frizzled/Smoothened family o frizzled o Smoothened

* Class B Secretin like o Calcitonin o Corticotropin releasing factor o Gastric inhibitory peptide o Glucagon o Growth hormone-releasing hormone o Parathyroid hormone o PACAP o Secretin o Vasoactive intestinal polypeptide o Diuretic hormone o EMR1 o Latrophilin o Brain-specific angiogenesis inhibitor (BAI) o Methuselah-like proteins (MTH) o Cadherin EGF LAG (CELSR) o Very large G-protein coupled receptor

Putative families: * Ocular albinism proteins * Insect odorant receptors * Plant Mlo receptors * Nematode chemoreceptors * Vomeronasal receptors (V1R & V3R) * Taste receptors T2R Orphans: * Putative / unclassified GPCRs non-GPCR families:

* Class Z Archaeal/bacterial/fungal opsins

GPCR Ligands
Rhodopsin family: amine receptors Acetylcholine (muscarinic) Adrenaline Dopamine Histamine Serotonin Octopamine Trace amine
Rhodopsin family: peptide receptors Angiotensin Apelin Bombesin Bradykinin C5a anaphylatoxin CC Chemokine CXC Chemokine CX3C Chemokine C Chemokine Cholecystokinin Endothelin fMet-Leu-Phe Galanin Ghrelin KiSS1-derived peptide Melanocortin Motilin Neuromedin U Neuropeptide FF Neuropeptide S Neuropeptide Y Neuropeptide W / neuropeptide B Neurotensin Orexigenic neuropeptide QRFP Opioid Orexin Oxytocin Prokineticin Somatostatin Tachykinin Urotensin II Vasopressin Protease-activated (thrombin) Adrenomedullin (G10D) GPR37 / endothelin B like Chemokine receptor like Melanin-concentrating hormone Follicle stimulating hormone Lutropin-choriogonadotropic hormone Thyrotropin

Rhodopsin family: other receptors Rhodopsin Olfactory Prostaglandin Prostacyclin Thromboxane Adenosine Purine / pyrimidine Cannabinoid Platelet activating factor Gonadotropin-releasing hormone Thyrotropin-releasing hormone

Metabotropic glutamate family Glutamate (metabotropic) Extracellular calcium-sensing GABA-B Pheromone (V2R) Taste receptors (T1R) Orphan GPRC5 Orphan GPCR6 Bride of sevenless proteins (BOSS) Putative / unclassified Class C GPCRs
Other families Frizzled / Smoothened family Ocular albinism proteins Vomeronasal receptors (V1R) Taste receptors (T2R) Insect odorant receptors Nematode chemoreceptors Plant Mlo receptors Fungal pheromone cAMP (Dictyostelium) Bacterial rhodopsin

Melatonin
Lysosphingolipid and LPA (EDG) Leukotriene B4 receptor SREB Mas proto-oncogene & Mas-related (MRGs) RDC1 EBV-induced Relaxin LGR like Free fatty acid G protein-coupled bile acid Nicotinic acid GPR GPR45 like Cysteinyl leukotriene Putative / unclassified Class A GPCRs Secretin family Calcitonin Corticotropin releasing factor Gastric inhibitory peptide Glucagon Growth hormone-releasing hormone Parathyroid hormone PACAP Secretin Vasoactive intestinal polypeptide EMR1 Latrophilin Brain-specific angiogenesis inhibitor (BAI) Methuselah-like proteins (MTH) Cadherin EGF LAG (CELSR) Putative / unclassified Class B GPCRs

G protein-based disease
GHRH
Pituitary GHRH Receptor

pituitary tumor

Gs
(+) cAMP (-)
Gi
GHRH--Growth-hormone-releasing hormone

GH secretion

somatostatin

GH--Growth-hormone

mechanism
Gs gene mutation GTPase activity Persistent activation of Gs

Persistent activation of AC
cAMP Pituitary proliferation and secretion Acromegaly or Gigantism

Cholera Toxin
cholera toxin enzyme that catalyzes the transfer of ADP ribose from intracellular NAD+ to alpha s. The ADP ribosylation alters the alpha s so that it can no longer hydrolyze its bound GTP. Thus, alpha s continues to stimulate adenylyl cyclase to produce cAMP. The prolonged production of cAMP in the intestinal epithelial cells causes a large efflux of Na+ and water into the gut, and is responsible for the severe diarrhea that is characteristic of cholera.

Effect of cholera toxin

Persistent activation of adenylyl cyclase

Reseptor Asetilkolin Muskarinik

Nestler et al, 2001 Molecular Neuropharmaclogy

Reseptor Asetilkolin Muskarinik

Reseptor Adrenergik

Receptors and signal transduction in the ANS

Adrenergic Receptors

1
1A 1B 1D 2A

2
2B 2C 1

Direct acting adrenergic receptor agonists: 1 receptors NH 3

Phenylephrine (Neosynephrine) Methoxamine (Vasoxyl) Oxymetazoline (Visine)
Phospho lipase C

(+)

Gq
PIP 2

HO
COOH IP 3 Increase Ca
2+

Diacylglycerol Activate Protein Kinase C

CH OH

C H2 N H

C H3

Ph en ylep h rin e
Response

Direct acting adrenergic receptor agonists: 2 receptors

Clonidine (Catapres) Methyldopa (Aldomet) Guanabenz (Wytensin) Guanfacine (Tenex) Tizanidine (Zanaflex)
NH 3

(-)

Adenylate Cyclase

Cl H N Cl Clo n id in e N N H

GI

K+

(+) ATP COOH

X
cAMP Reduce cAMP -Dependent Protein Kinase Activity

Response

Reseptor Dopamin

Nestler et al, 2001 Molecular Neuropharmaclogy,

OBAT YANG BEKERJA PADA SISTEM DOPAMINERGIK