G Protein-Coupled Receptors: Structure, Signaling, and Physiology

Half-title......Page 3
Title......Page 5
Copyright......Page 6
Contents......Page 7
Figures......Page 9
Tables......Page 13
Introduction......Page 19
Introduction......Page 23
The origin of GPCR genes......Page 24
Expansion of GPCR genes......Page 27
The loss of GPCR functions......Page 29
Structural shaping of the core of GPCRs......Page 31
Structural evolution of intra- and extracellular domains of GPCRs......Page 33
Selection on GPCR genes......Page 35
Genetic signatures of selection......Page 36
Selection of genomic regions containing GPCR genes......Page 37
Selection of individual GPCRs......Page 39
In vitro evolution of GPCRs......Page 40
Suggested readIng......Page 42
Introduction......Page 50
Structural and biophysical characterization of reconstituted lipoprotein particles......Page 51
Reconstitution of GPCRs into lipoprotein particles......Page 53
Isolation and analysis of monomeric and dimeric forms of GPCRs  in lipoprotein particles......Page 57
G protein coupling to GPCRs reconstituted in lipoprotein particles......Page 58
Conformational changes conferred by ligand binding 
and G protein coupling......Page 62
Conclusions......Page 66
References......Page 67
GPCRs and drug discovery......Page 71
GPCR dimerization......Page 72
GPCR-Gα protein fusions......Page 73
Applications to GPCR dimerization......Page 74
Heteromer-selective ligands......Page 78
Suggested readIng......Page 79
4 Time-resolved FRET approaches to study GPCR complexes......Page 85
Introduction......Page 86
Basic principle of FRET and main parameters: distance and orientation......Page 87
Limitations of FRET using conventional fluorophores......Page 88
Lanthanide-based fluorophores......Page 90
Acceptor fluorophores for TR-FRET......Page 93
Antibody-based TR-FRET approach validates GPCR dimers in transfected cells......Page 94
Limitation on the use of antibodies-based TR-FRET approach......Page 96
Covalent labeling of proteins with TR-FRET 
compatible fluorophores......Page 97
The snap-tag technology......Page 98
The ACP-tag technology......Page 99
Validation using class C GPCRs......Page 100
Class C GPCR oligomerization determined using snap-tag labeling......Page 102
Conclusion and perspectives......Page 103
References......Page 104
5 Signaling of dopamine receptor homo- and heterooligomers......Page 108
Dopamine receptors......Page 109
Dopamine receptor homooligomerization......Page 110
Heterooligomerization between adenosine A2A receptor and dopamine D2 receptor......Page 111
Heterooligomerization between µ-opioid receptor and dopamine D1 receptor......Page 112
Heterooligomerization of GABAergic receptor with dopamine D5 receptor......Page 113
Heterooligomerization of NMDA and D1 receptors......Page 114
Heterooligomerization of somatostatin SSTR5 and dopamine D2 receptor......Page 115
Interaction between D2 and D5 receptors......Page 116
Heterooligomerization of D1-D2 receptors......Page 117
The existence of D1-D2 receptor heterooligomers 
in the brain......Page 118
Physiological relevance......Page 119
Suggested readIng......Page 121
Introduction......Page 129
Chemokines......Page 130
Chemokine receptors......Page 131
Methods to determine receptor dimerization......Page 132
Dynamics of chemokine receptor dimers......Page 134
Therapeutic opportunities of chemokine receptor dimerization......Page 135
Concluding remarks......Page 136
References......Page 137
Introduction......Page 143
Development......Page 144
Central nervous system......Page 148
Hematopoiesis and immune system......Page 150
Cardiovascular system......Page 151
Endocrine system and metabolism......Page 153
Sensory systems......Page 154
Hemostasis......Page 155
Suggested readIng......Page 156
Introduction......Page 163
Ligand binding......Page 164
Receptor activation......Page 165
Receptor/G protein interaction......Page 167
G protein activation......Page 168
Effector molecules and second messengers......Page 169
Interlude – where is the time-limiting step?......Page 170
GPCR phosphorylation and arrestin binding......Page 171
EpIlogue......Page 172
Suggested readIng......Page 173
9 RGS-RhoGEFs and other RGS multidomain proteins as effector molecules in GPCR-dependent and GPCR-independent cell signaling......Page 177
Introduction......Page 178
p115RhoGEF, PDZ-RhoGEF, and LARG......Page 180
Cloning of Gα12/13-regulated RGS-RhoGEFs......Page 182
p115-RhoGEF.......Page 183
LARG. ......Page 184
Thrombin and lysophosphatidic acid receptors as paradigms 
of GPCR signaling to Rho through RGS-RhoGEFs......Page 185
Phosphorylation......Page 186
Oligomerization......Page 187
Role in Plexin-B Signaling......Page 189
Role in tyrosine kinase receptor signaling......Page 191
Role in spatiotemporal control of RhoGTPases, actin and microtubule dynamics, cell migration and polarity......Page 192
Structural basis of GPCR signaling to RGS-RhoGEFs......Page 188
RGS-RhoGEFs in the nervous system......Page 193
RGS-RhoGEFs in the immune system......Page 194
Potential role in diabetes......Page 195
Gq signaling to Rho......Page 196
Gq/GRK2......Page 197
G Proteins/Axin......Page 198
Gi/RGS12, RGS14......Page 199
RGS12 as a scaffold in NGF-mediated differentiation......Page 200
Acknowledgments......Page 201
References......Page 202
10 Adenylyl cyclase isoform-specific signaling of GPCRs......Page 207
Cyclic AMP signaling pathway......Page 208
Expression of adenylyl cyclases......Page 209
Historical perspective of GPCR modulation of adenylyl cyclase......Page 210
GPCR regulation of group I adenylyl cyclases (AC1, AC3, and AC8)......Page 213
GPCR regulation of group II adenylyl cyclases (AC2, AC4, and AC7)......Page 214
GPCR regulation of group III adenylyl cyclases (AC5 and AC6)......Page 215
GPCR regulation of group IV adenylyl cyclases (AC9)......Page 216
Heterologous sensitization......Page 217
Role of Gβγ in heterologous sensitization of adenylyl cyclase......Page 218
Gα.s. and adenylyl cyclase isoform differences in heterologous sensitization......Page 219
Posttranslational modifications and GPCR signaling......Page 220
Regulation of cyclic AMP signaling by oligomerization of adenylyl cyclases......Page 221
Compartmentalization of adenylyl cyclases and GPCRs in membrane rafts......Page 222
Direct interactions between GPCR and adenylyl cyclase isoforms......Page 223
Impact of GPCR heterooligomers on adenylyl cyclase signaling......Page 224
Effects of Gβ and Gγ modulation on GPCR-adenylyl cyclase signaling......Page 225
Adenylyl cyclase knockout mice......Page 226
Concluding Remarks......Page 227
References......Page 228
Introduction......Page 235
Receptor phosphorylation......Page 236
Receptor desensitization......Page 237
Receptor endocytosis......Page 238
ERK MAP kinases......Page 239
DGK......Page 241
Phosphorylation......Page 242
Ubiquitinylylation......Page 243
Cell proliferation......Page 244
References......Page 245
GPCR ligands in drug discovery......Page 249
GPCR ligand binding......Page 251
GPCR downstream signaling......Page 253
High content screening (HCS)......Page 254
HCS to measure GPCR ligand binding, internalization, and arrestin signaling......Page 255
HCS to measure various GPCR downstream signaling events......Page 257
Discussion......Page 259
Suggested readIng......Page 260
Introduction......Page 265
The nature of receptor allosterism......Page 266
Effects of β, the intrinsic efficacy of the allosteric modulator......Page 270
Effect of γ, the binding cooperativity of the allosteric modulator......Page 271
Examples of allosteric modulators......Page 272
Potential (therapeutic) advantages 
of allosteric modulators......Page 278
Summary and Conclusions......Page 280
Suggested readIng......Page 281
Introduction: 7TM receptors......Page 288
Functional selectivity: historical context......Page 289
Thermodynamic mechanism(s)......Page 292
Pharmacological assays......Page 295
A quantitative scale for functional selectivity......Page 296
Conclusions......Page 298
References......Page 299
15 β -adrenoceptors in cardiovascular and respiratory diseases......Page 305
βAR Structure and Subtypes......Page 306
βAR activation and signaling......Page 308
Role of βARs in regulation of cardiovascular and respiratory function......Page 309
Cardiovascular diseases......Page 310
Respiratory diseases......Page 311
Hypertension......Page 312
βAR antagonists in acute and chronic myocardial ischemia......Page 313
Applications of β-blockers in heart failure and cardiomyopathy......Page 314
Use of β-blockers for arrhythmias......Page 315
GRKs and cardiac diseases......Page 316
GRKs in respiratory diseases......Page 317
Human β1AR polymorphisms......Page 318
Human β2AR polymorphisms......Page 320
Risk of developing heart failure and therapeutic responses in cardiac diseases with βAR SNPs......Page 322
Pharmacogenetic interaction in heart failure patients between β-blockers and βAR SNPs......Page 323
Effects of β2AR SNPs on asthma therapy......Page 324
References......Page 325
16 Role of metabotropic glutamate receptors in CNS disorders......Page 339
Glutamate in the brain......Page 340
Group I Mglurs......Page 343
Group I Mglurs in anxiety disorders......Page 344
Group I Mglurs in cognitive disorders......Page 345
Group I Mglurs in schizophrenia......Page 347
Fragile X syndrome......Page 351
Group I Mglurs in Parkinson’s disease......Page 353
Group I Mglurs in epilepsy......Page 356
Group I Mglurs in pain disorders......Page 358
Group II Mglurs......Page 359
Group II Mglurs in anxiety disorders......Page 362
Group II Mglurs in cognitive disorders......Page 364
Group II Mglurs in schizophrenia......Page 366
Group II Mglurs in addiction......Page 367
Depression......Page 369
Group II Mglurs in epilepsy......Page 370
Group II mGluRs in pain disorders......Page 371
Group III Mglurs......Page 374
Group III Mglurs in anxiety disorders......Page 375
Group III Mglurs in schizophrenia......Page 377
Group III Mglurs in Parkinson’s disease......Page 378
Group III Mglurs in pain disorders......Page 379
Group III Mglurs in retinal dysfunction......Page 381
Conclusions and perspectives......Page 382
Sphingosine 1-phosphate......Page 398
Sphingolipid rheostat and sphingosine kinases......Page 399
S1P gradient and cellular sources of S1P......Page 400
Sphingosine 1-phosphate receptors......Page 401
S1P1 receptor......Page 402
S1P2 and S1P3......Page 404
S1P receptors in immune function......Page 405
Mechanism of lymphopenia evoked by S1P1 agonists......Page 406
Effect of S1P on Th17 cell generation......Page 407
Fingolimod: Clinical validation of S1P receptor agonism in multiple sclerosis......Page 408
Novel therapeutic approaches targeting S1P system......Page 409
Concluding remarks......Page 410
References......Page 411
Introduction to Wnt signaling......Page 416
Frizzled receptors: central role in Wnt signaling......Page 417
Canonical Wnt signaling......Page 418
Planar cell polarity signaling......Page 420
Ca.2+. signaling......Page 421
Wnt/Frizzled pathway implication in bone metabolism......Page 422
Genetic models......Page 424
Translating Wnt into therapeutics......Page 425
Frizzled receptors as potential drug target for treatment of bone diseases......Page 426
References......Page 427
Index......Page 433