Graduate Institute of Anatomy and Cell Biology

G-protein-coupled receptors
(GPCRs)

mchuang@ntu.edu.tw
Graduate Institute of Anatomy and Cell Biology

The Nobel Prize in Chemistry 2012 is awarded to Brian K. Kobilka (57)

and Robert J. Lefkowitz (69) for studies of GPCRs.

 &

G-protein-coupled receptors (GPCRs)

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G protein-coupled receptors (GPCRs)

Seven transmembrane receptors (7 TM receptors)
Heptahelical receptors
Serpentine receptors
CCR5

Single Transmembrane Spanning Heterotrimeric G

Protein-Coupled Receptors

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Pharmacol Rev. 2004 Sep;56(3):371-85.

Characteristics of GPCRs
Extracellular

NH2

7 transmembrane domains
3 intracellular loops

Cytoplasmic

3 extracellular loops

HOOC
A Transmembrane Intracellular Estrogen Receptor Mediates Rapid Cell Signaling.
GPR30 is uniquely localized to the endoplasmic reticulum
Revankar CM et al., Science. 2005. 307:1625.
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G-protein-coupled receptors (GPCRs)

Genomics: > 800 members

wide range of cellular
activities
from yeast to human
50% drug target


G-protein-coupled receptors (GPCRs)
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G-protein-coupled receptor (GPCR)

families

Class A

Rhodopsin like

Class B

Secretin like

Class C

Metabotropic glutamate / pheromone

Class D

Fungal pheromone

Class E

cAMP receptors (Dictyostelium)

Frizzled/Smoothened family
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Family classification of known GPCRs

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The three main families of GPCRs

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GRAFS system
Five main classes: Glutamate, Rhodopsin, Adhesion, Frizzled, and Secretin
Orphan receptors
(unknown ligands)

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Annu Rev Pharmacol Toxicol. 2013;53:127-46

G-protein-coupled receptors (GPCRs)

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http://stke.sciencemag.org/content/vol0/issue2002/images/data/CMP_6634/DC1/science3.jpg, by R. Iyengar's laboratory

Summary of GPCR signalling

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NATURE | VOL 415 | 10 JANUARY 2002 206-212

G protein action
ligand

1.
GPCR

2.
GPCR
(GEF)

GTP

GDP

GDP

3.

4.

effector
GPCR

GDP

Pi

GPCR

stops

(GAP)

GTP
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second messenger

G protein cycle for the 2ARGs complex

Kobilkas group
Nature. 2011 Jul 19;477(7366):549-55.

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A classical example of GPCR signalling

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4
GPCR

G protein

effector

activities

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Modified from TRENDS in Pharmacological Sciences Vol.22 No.7 July 2001, 368-376

Regulation of systemic functions by signaling through

G protein pathways

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31 MAY 2002 VOL 296 SCIENCE, 1636-1639

Heterotrimeric G-protein -subunit subfamilies

and their effects

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Desensitization

G-protein-coupled receptors (GPCRs)

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Desensitization
(receptor level)
(A) Rapid: phosphorylation (sec~min)
(B) Slow: degradation or expression (hours)

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Desensitization
(A) Rapid: phosphorylation (sec~min)
1. Second-messenger kinase (PKA, PKC):
uncouple GPCR from G protein
ligand independent
heterologous desensitization

2. GPCR kinase (GRK): GRK1-GRK7

binding of -arrestin and inhibition of G protein interaction

ligand-activated or activated conformation
homologous desensitization
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Desensitization
GPCR kinase (GRK) phosphorylation
arrestin 1-arrestin 4

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Desensitization: (B) Degradation

Recycling or degradation?

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SCIENCE VOL 297 26 JULY 2002

Desensitization
(Post-receptor level)
RGS (regulator of G-protein signaling)
a. GTPase-activating proteins
b. accelerate GTP hydrolysis
c. >25 members, for selectivity
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Desensitization by
regulator of G-protein signalling (RGS)

RGS2 knockout mice:

T-cell activation
Anxiety
Male aggression
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Internalization

G-protein-coupled receptors (GPCRs)

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Internalization
Functions:
1. Desensitization
2. Resensitization
3. Signal transduction
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Internalization
Types:
Clathrin-coated pits
-arrestin

Caveolae-mediated
Lipid rafts: signalling

Uncoated vesicles
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Function 1: Desensitization
-arrestin-dependent internalization of GPCRs

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NATURE REVIEWS | NEUROSCIENCE VOLUME 2 | OCTOBER 2001 | 727-733

Function 2: Resensitization

GRP: GPCR specific

phosphatase

P
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Pharmacol Rev 53:124, 2001

Function 3: Signal transduction

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Annu. Rev. Physiol. 2007. 69:51134

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TRENDS in Pharmacological Sciences Vol.28 No.8 417

GPCR signaling via heterotrimeric G proteins from endosomes

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J Biol Chem. 2015 Jan 20. [Epub ahead of print]

G-protein-independent
signalling

G-protein-coupled receptors (GPCRs)

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Regulation of GPCR Signalling

by Scaffold Proteins

G-protein-independent signalling
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Scaffold Proteins

3
2

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1. GPCR/PDZ scaffold interaction

(calcium channel)

PDZ: PSD-95/Discs-large/ZO-1
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Circ Res. 2002;91:672-680

2 adrenoceptor regulates the Na+-H+ exchanger

(NHE)

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TRENDS in Biochemical Sciences Vol.26 No.5 May 2001, 291-297

2. GPCR/non-PDZ scaffold interaction

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Circ Res. 2002;91:672-680

Metabotropic glutamate receptor-mediated Ca2+ release

Ca2+ release

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TRENDS in Biochemical Sciences Vol.26 No.5 May 2001, 291-297

3. GPCR/-arrestin interaction

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Circ Res. 2002;91:672-680

2 Adrenoceptor can mediate the Src kinase

pathway

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TRENDS in Biochemical Sciences Vol.26 No.5 May 2001, 291-297

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Transactivation

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Trends Endocrinol Metab. 2006 Mar;17(2):48-54

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Trends Endocrinol Metab. 2006 Mar;17(2):48-54

Oligomerization

G-protein-coupled receptors (GPCRs)

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Signal amplification at the receptor level

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Heterodimerization of the GABAB receptor produces

a functional receptor

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Dont worry B happy!:

a role for GABAB
receptors in anxiety and
depression
John F. Cryan and Klemens
Kaupmann
Neuroscience Research, Novartis
Institutes for BioMedical Research,
Novartis Pharma AG, Basel,
Switzerland

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TRENDS in Pharmacological Sciences Vol.26 No.1 January 2005

Ligand-independent functions of orphan GPCRs in heterodimers

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EMBO reports VOL 7 | NO 11 | 2006, p. 1096

Bivalent and dimeric ligands

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Drug discovery
GPCR Drugs >$20B annual sales

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GPCR and Diseases

Obesity
Cardiovascular Disease
Inflammation
Cancer
Diabetes
Alzheimers Disaese
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Drug screening

G-protein-coupled receptors (GPCRs)

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Key requirements of a ligand screen

Cell surface expression of GPCRs
Cellular assays to detect ligand binding
Libraries of putative natural and
synthetic ligands
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Method for de-orphanizing GPCRs

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FIRST : Functionally Immobilized Receptor

Solubilization Technology

Consensus Pharmaceuticals, Inc.

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CART
Constitutively Active Receptor Technology

Arena Pharmaceuticals, Inc.

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Diseases - GPCRs
DISEASE
OVERACTIVE KNOWN GPCRs
---------------------------------------------------------------Schizophrenia
5HT2A, D4
---------------------------------------------------------------Depression
5HT2A
---------------------------------------------------------------Hyperthyroidism
Thyrotropin
---------------------------------------------------------------Hypertension
Angiotensin AT1A
---------------------------------------------------------------Asthma
Adenosine A1
---------------------------------------------------------------Melanoma
MC-1
---------------------------------------------------------------Retinitis Pigmentosa
Rhodopsin
---------------------------------------------------------------63

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Nat Rev Drug Discov. 2011 Jan;10(1):47-60.

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Nat Rev Drug Discov. 2011 Jan;10(1):47-60.

Representation of inactive and active GPCR conformations

inactive

active
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NATURE REVIEWS | DRUG DISCOVERY VOLUME 1 | AUGUST 2002 | 599-608

CART
Constitutively Active Receptor Technology
Inverse agonists: block ligand-independent activity
Selective molecules: one ligand to multiple receptors
Agonists
Allosteric modulators
(Antagonist: block ligand-dependent activity)
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Summary

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Diversity of G-protein-coupled receptors (GPCRs)

Internalization

Scaffold proteins

GRK
PKA
PKC

signaling

RGS

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Modified from TRENDS in Pharmacological Sciences Vol.22 No.7 July 2001, 368-376

Treasures throughout the life-cycle of GPCRs

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TRENDS in Pharmacological Sciences Vol.22 No.8 August 2001, 396