Identification of the endophilins (SH3p4/p8/p13) as novel binding partners for the beta1-adrenergic receptor.

Tang, Y

Hu, LA

Miller, WE

Ringstad, N

Hall, RA

Pitcher, JA

DeCamilli, P

Lefkowitz, RJ

United States

2013-09-05T18:13:44Z

1999-10-26

Several G-protein coupled receptors, such as the beta1-adrenergic receptor (beta1-AR), contain polyproline motifs within their intracellular domains. Such motifs in other proteins are known to mediate protein-protein interactions such as with Src homology (SH)3 domains. Accordingly, we used the proline-rich third intracellular loop of the beta1-AR either as a glutathione S-transferase fusion protein in biochemical "pull-down" assays or as bait in the yeast two-hybrid system to search for interacting proteins. Both approaches identified SH3p4/p8/p13 (also referred to as endophilin 1/2/3), a SH3 domain-containing protein family, as binding partners for the beta1-AR. In vitro and in human embryonic kidney (HEK) 293 cells, SH3p4 specifically binds to the third intracellular loop of the beta1-AR but not to that of the beta2-AR. Moreover, this interaction is mediated by the C-terminal SH3 domain of SH3p4. Functionally, overexpression of SH3p4 promotes agonist-induced internalization and modestly decreases the Gs coupling efficacy of beta1-ARs in HEK293 cells while having no effect on beta2-ARs. Thus, our studies demonstrate a role of the SH3p4/p8/p13 protein family in beta1-AR signaling and suggest that interaction between proline-rich motifs and SH3-containing proteins may represent a previously underappreciated aspect of G-protein coupled receptor signaling.

https://www.ncbi.nlm.nih.gov/pubmed/10535961

0027-8424

https://hdl.handle.net/10161/7817

eng

Proceedings of the National Academy of Sciences

Proc Natl Acad Sci U S A

Adaptor Proteins, Signal Transducing

Adrenergic beta-1 Receptor Agonists

Animals

Carrier Proteins

Cattle

Cell Line

GTP-Binding Protein alpha Subunits, Gs

Humans

Proline

Protein Binding

Receptors, Adrenergic, beta-1

src Homology Domains

Identification of the endophilins (SH3p4/p8/p13) as novel binding partners for the beta1-adrenergic receptor.

Journal article

https://www.ncbi.nlm.nih.gov/pubmed/10535961

12559

12564

22

Basic Science Departments

Biochemistry

Chemistry

Clinical Science Departments

Duke

Duke Cancer Institute

Institutes and Centers

Medicine

Medicine, Cardiology

Pathology

School of Medicine

Trinity College of Arts & Sciences

Published

96