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Spinophilin Blocks Arrestin Actions in Vitro and in Vivo at G Protein-Coupled Receptors
Qin Wang,1
Jiali Zhao,1
Ashley E. Brady,1
Jian Feng,2
Patrick B. Allen,3
Robert J. Lefkowitz,4
Paul Greengard,5
Lee E. Limbird1*
Abstract:
Arrestin regulates almost all G protein–coupled receptor(GPCR)–mediated signaling and trafficking. We report thatthe multidomain protein, spinophilin, antagonizes these multiplearrestin functions. Through blocking G protein receptor kinase2 (GRK2) association with receptor-Gß complexes, spinophilinreduces arrestin-stabilized receptor phosphorylation, receptorendocytosis, and the acceleration of mitogen-activated proteinkinase (MAPK) activity following endocytosis. Spinophilin knockoutmice were more sensitive than wild-type mice to sedation elicitedby stimulation of 2 adrenergic receptors, whereas arrestin 3knockout mice were more resistant, indicating that the signal-promoting,rather than the signal-terminating, roles of arrestin are moreimportant for certain response pathways. The reciprocal interactionsof GPCRs with spinophilin and arrestin represent a regulatorymechanism for fine-tuning complex receptor-orchestrated cellsignaling and responses.
1 Department of Pharmacology and Center of Molecular Neuroscience, Vanderbilt University Medical Center, Nashville, TN 37232, USA. 2 Department of Physiology and Biophysics, State University of New York at Buffalo, Buffalo, NY 14214, USA. 3 Department of Psychiatry, Yale University School of Medicine, New Haven, CT 06508, USA. 4 Howard Hughes Medical Institute, Department of Medicine and Biochemistry, Duke University Medical Center, Durham, NC 27708, USA. 5 Laboratory of Molecular and Cellular Neuroscience, The Rockefeller University, New York, NY 10021, USA.
* To whom correspondence should be addressed at Department of Pharmacology, Vanderbilt University Medical Center, 464 Robinson Research Building, Nashville, TN 37232–6600, USA. E-mail: lee.limbird{at}mcmail.vanderbilt.edu
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complexes, spinophilin reduces arrestin-stabilized receptor phosphorylation, receptor endocytosis, and the acceleration of mitogen-activated protein kinase (MAPK) activity following endocytosis. Spinophilin knockout mice were more sensitive than wild-type mice to sedation elicited by stimulation of 
2 adrenergic receptors, whereas arrestin 3 knockout mice were more resistant, indicating that the signal-promoting, rather than the signal-terminating, roles of arrestin are more important for certain response pathways. The reciprocal interactions of GPCRs with spinophilin and arrestin represent a regulatory mechanism for fine-tuning complex receptor-orchestrated cell signaling and responses. 
