Regulation of beta-adrenergic receptor signaling by S-nitrosylation of G-protein-coupled receptor kinase 2.
beta-adrenergic receptors (beta-ARs), prototypic G-protein-coupled receptors (GPCRs), play a critical role in regulating numerous physiological processes. The GPCR kinases (GRKs) curtail G-protein signaling and target receptors for internalization. Nitric oxide (NO) and/or S-nitrosothiols (SNOs) can prevent the loss of beta-AR signaling in vivo, but the molecular details are unknown. Here we show in mice that SNOs increase beta-AR expression and prevent agonist-stimulated receptor downregulation; and in cells, SNOs decrease GRK2-mediated beta-AR phosphorylation and subsequent recruitment of beta-arrestin to the receptor, resulting in the attenuation of receptor desensitization and internalization. In both cells and tissues, GRK2 is S-nitrosylated by SNOs as well as by NO synthases, and GRK2 S-nitrosylation increases following stimulation of multiple GPCRs with agonists. Cys340 of GRK2 is identified as a principal locus of inhibition by S-nitrosylation. Our studies thus reveal a central molecular mechanism through which GPCR signaling is regulated.
Whalen
Erin J
EJ
Department of Medicine, Duke University Medical Center, Durham, NC 27710, USA.
Foster
Matthew W
MW
Matsumoto
Akio
A
Ozawa
Kentaro
K
Violin
Jonathan D
JD
Que
Loretta G
LG
Nelson
Chris D
CD
Benhar
Moran
M
Keys
Janelle R
JR
Rockman
Howard A
HA
Koch
Walter J
WJ
Daaka
Yehia
Y
Lefkowitz
Robert J
RJ
Stamler
Jonathan S
JS
D000818:Animals; D019390:Arrestins / Q000378:metabolism; D002460:Cell Line; D045744:Cell Line, Tumor; D003545:Cysteine / Q000378:metabolism; D054769:G-Protein-Coupled Receptor Kinase 2; D006706:Homeostasis; D006801:Humans; D008168:Lung / Q000378:metabolism; D051379:Mice; D009206:Myocardium / Q000378:metabolism; D017942:Nitric Acid / Q000378:metabolism; D010766:Phosphorylation; D011943:Receptors, Adrenergic, beta / Q000378:metabolism*; D026403:S-Nitrosothiols / Q000378:metabolism*; D015398:Signal Transduction; D051552:beta-Adrenergic Receptor Kinases / Q000737:chemistry / Q000378:metabolism*; D000071557:beta-Arrestins
D019390:Arrestins; D011943:Receptors, Adrenergic, beta; D026403:S-Nitrosothiols; D000071557:beta-Arrestins; D017942:Nitric Acid; C496567:GRK2 protein, human; C496568:GRK2 protein, mouse; D051552:beta-Adrenergic Receptor Kinases; D054769:G-Protein-Coupled Receptor Kinase 2; D003545:Cysteine