Note to users. If you're seeing this message, it means that your browser cannot find this page's style/presentation instructions -- or possibly that you are using a browser that does not support current Web standards. Find out more about why this message is appearing, and what you can do to make your experience of our site the best it can be.
Career Basics

Site Tools

  • AAAS
  • Subscribe
  • Feedback

Site Search

Search Advanced

Logo for

PNAS 96 (22): 12559-12564

Copyright © 1999 by the National Academy of Sciences.

Vol. 96, Issue 22, 12559-12564, October 26, 1999

Cell Biology
 Identification of the endophilins (SH3p4/p8/p13) as novel binding partners for the beta 1-adrenergic receptor

Yuting Tang*,dagger , Liaoyuan A. Hu*,dagger , William E. Miller*,dagger , Niels RingstadDagger , Randy A. Halldagger ,§, Julie A. Pitcherdagger ,, Pietro DeCamilliDagger , and Robert J. Lefkowitz*,dagger ,parallel

* Howard Hughes Medical Institute and dagger  Department of Medicine (Cardiology) and Biochemistry, P.O. Box 3821, Duke University Medical Center, Durham, NC 27710; and Dagger  Yale University School of Medicine, New Haven, CT 06150

Contributed by Robert J. Lefkowitz, September 1, 1999

Several G-protein coupled receptors, such as the beta 1-adrenergic receptor (beta 1-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 beta 1-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 beta 1-AR. In vitro and in human embryonic kidney (HEK) 293 cells, SH3p4 specifically binds to the third intracellular loop of the beta 1-AR but not to that of the beta 2-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 beta 1-ARs in HEK293 cells while having no effect on beta 2-ARs. Thus, our studies demonstrate a role of the SH3p4/p8/p13 protein family in beta 1-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.

§   Present address: Department of Pharmacology, Rollins Research Center, Emory University School of Medicine, Atlanta, GA 30322.
   Present address: Medical Research Council Laboratory for Molecular Cell Biology, University College London, Gower Street, London, United Kingdom, WC1E 6BT.
parallel    To whom reprint requests should be addressed. E-mail: lefko001{at}mc.duke.edu.

Copyright © 1999 by The National Academy of Sciences  0027-8424/99/9612559-6$2.00/0

THIS ARTICLE HAS BEEN CITED BY OTHER ARTICLES:
eena Promotes Myeloid Proliferation through Stimulating ERK1/2 Phosphorylation in Zebrafish.
H.-Y. Le, Y. Zhang, H. Liu, L.-H. Ma, Y. Jin, Q.-H. Huang, Y. Chen, M. Deng, Z. Chen, S.-J. Chen, et al. (2008)
J. Biol. Chem. 283, 17652-17661
   Abstract »    Full Text »    PDF »
Endocytosis machinery is required for {beta}1-adrenergic receptor-induced hypertrophy in neonatal rat cardiac myocytes.
C. Morisco, C. Marrone, J. Galeotti, D. Shao, D. E. Vatner, S. F. Vatner, and J. Sadoshima (2008)
Cardiovasc Res 78, 36-44
   Abstract »    Full Text »    PDF »
Aberrant transcriptional regulation of the MLL fusion partner EEN by AML1-ETO and its implication in leukemogenesis.
L.-H. Ma, H. Liu, H. Xiong, B. Chen, X.-W. Zhang, Y.-Y. Wang, H.-Y. Le, Q.-H. Huang, Q.-H. Zhang, B.-L. Li, et al. (2007)
Blood 109, 769-777
   Abstract »    Full Text »    PDF »
Two synaptojanin 1 isoforms are recruited to clathrin-coated pits at different stages.
R. M. Perera, R. Zoncu, L. Lucast, P. De Camilli, and D. Toomre (2006)
PNAS 103, 19332-19337
   Abstract »    Full Text »    PDF »
AKAP79-mediated Targeting of the Cyclic AMP-dependent Protein Kinase to the beta1-Adrenergic Receptor Promotes Recycling and Functional Resensitization of the Receptor.
L. A. Gardner, S. J. Tavalin, A. S. Goehring, J. D. Scott, and S. W. Bahouth (2006)
J. Biol. Chem. 281, 33537-33553
   Abstract »    Full Text »    PDF »
Association of 14-3-3 Proteins to beta1-Adrenergic Receptors Modulates Kv11.1 K+ Channel Activity in Recombinant Systems.
A. S. Tutor, E. Delpon, R. Caballero, R. Gomez, L. Nunez, M. Vaquero, J. Tamargo, F. Mayor Jr., and P. Penela (2006)
Mol. Biol. Cell 17, 4666-4674
   Abstract »    Full Text »    PDF »
Trafficking of G Protein-Coupled Receptors.
M. T. Drake, S. K. Shenoy, and R. J. Lefkowitz (2006)
Circ. Res. 99, 570-582
   Abstract »    Full Text »    PDF »
The BAR Domain Proteins: Molding Membranes in Fission, Fusion, and Phagy.
G. Ren, P. Vajjhala, J. S. Lee, B. Winsor, and A. L. Munn (2006)
Microbiol. Mol. Biol. Rev. 70, 37-120
   Abstract »    Full Text »    PDF »
Proteomic Analysis of beta1-Adrenergic Receptor Interactions with PDZ Scaffold Proteins.
J. He, M. Bellini, H. Inuzuka, J. Xu, Y. Xiong, X. Yang, A. M. Castleberry, and R. A. Hall (2006)
J. Biol. Chem. 281, 2820-2827
   Abstract »    Full Text »    PDF »
Agonist-independent Activation of Src Tyrosine Kinase by a Cholecystokinin-2 (CCK2) Receptor Splice Variant.
B. Olszewska-Pazdrak, C. M. Townsend Jr., and M. R. Hellmich (2004)
J. Biol. Chem. 279, 40400-40404
   Abstract »    Full Text »    PDF »
RGS2 Binds Directly and Selectively to the M1 Muscarinic Acetylcholine Receptor Third Intracellular Loop to Modulate Gq/11{alpha} Signaling.
L. S. Bernstein, S. Ramineni, C. Hague, W. Cladman, P. Chidiac, A. I. Levey, and J. R. Hepler (2004)
J. Biol. Chem. 279, 21248-21256
   Abstract »    Full Text »    PDF »
Src Homology 3 Binding Sites in the P2Y2 Nucleotide Receptor Interact with Src and Regulate Activities of Src, Proline-rich Tyrosine Kinase 2, and Growth Factor Receptors.
J. Liu, Z. Liao, J. Camden, K. D. Griffin, R. C. Garrad, L. I. Santiago-Perez, F. A. Gonzalez, C. I. Seye, G. A. Weisman, and L. Erb (2004)
J. Biol. Chem. 279, 8212-8218
   Abstract »    Full Text »    PDF »
Identification and characterization of EBP, a novel EEN binding protein that inhibits Ras signaling and is recruited into the nucleus by the MLL-EEN fusion protein.
J. W. P. Yam, D.-Y. Jin, C. W. So, and L. C. Chan (2004)
Blood 103, 1445-1453
   Abstract »    Full Text »    PDF »
The Angiotensin II Type I Receptor-associated Protein, ATRAP, Is a Transmembrane Protein and a Modulator of Angiotensin II Signaling.
M. Lopez-Ilasaca, X. Liu, K. Tamura, and V. J. Dzau (2003)
Mol. Biol. Cell 14, 5038-5050
   Abstract »    Full Text »    PDF »
Resistance of the Human {beta}1-Adrenergic Receptor to Agonist-mediated Down-regulation: ROLE OF THE C TERMINUS IN DETERMINING {beta}-SUBTYPE DEGRADATION.
W. Liang, S. Austin, Q. Hoang, and P. H. Fishman (2003)
J. Biol. Chem. 278, 39773-39781
   Abstract »    Full Text »    PDF »
Mutational Uncoupling of {alpha}1A-Adrenergic Receptors from G Proteins Also Uncouples Mitogenic and Transcriptional Responses in PC12 Cells.
D. Lee, A. Robeva, Z. Chen, and K. P. Minneman (2003)
J. Pharmacol. Exp. Ther. 306, 471-477
   Abstract »    Full Text »    PDF »
GIPC Interacts with the {beta}1-Adrenergic Receptor and Regulates {beta}1-Adrenergic Receptor-mediated ERK Activation.
L. A. Hu, W. Chen, N. P. Martin, E. J. Whalen, R. T. Premont, and R. J. Lefkowitz (2003)
J. Biol. Chem. 278, 26295-26301
   Abstract »    Full Text »    PDF »
G-protein-coupled Receptor (GPCR) Kinase Phosphorylation and {beta}-Arrestin Recruitment Regulate the Constitutive Signaling Activity of the Human Cytomegalovirus US28 GPCR.
W. E. Miller, D. A. Houtz, C. D. Nelson, P. E. Kolattukudy, and R. J. Lefkowitz (2003)
J. Biol. Chem. 278, 21663-21671
   Abstract »    Full Text »    PDF »
Characterization of Endophilin B1b, a Brain-specific Membrane-associated Lysophosphatidic Acid Acyl Transferase with Properties Distinct from Endophilin A1.
J. Modregger, A. A. Schmidt, B. Ritter, W. B. Huttner, and M. Plomann (2003)
J. Biol. Chem. 278, 4160-4167
   Abstract »    Full Text »    PDF »
Mitral Cell beta 1 and 5-HT2A Receptor Colocalization and cAMP Coregulation: A New Model of Norepinephrine-Induced Learning in the Olfactory Bulb.
Q. Yuan, C. W. Harley, and J. H. McLean (2003)
Learn. Mem. 10, 5-15
   Abstract »    Full Text »    PDF »
Alix (ALG-2-interacting Protein X), a Protein Involved in Apoptosis, Binds to Endophilins and Induces Cytoplasmic Vacuolization.
C. Chatellard-Causse, B. Blot, N. Cristina, S. Torch, M. Missotten, and R. Sadoul (2002)
J. Biol. Chem. 277, 29108-29115
   Abstract »    Full Text »    PDF »
Novel Interaction between the M4 Muscarinic Acetylcholine Receptor and Elongation Factor 1A2.
D. B. McClatchy, C. R. Knudsen, B. F. Clark, R. A. Kahn, R. A. Hall, and A. I. Levey (2002)
J. Biol. Chem. 277, 29268-29274
   Abstract »    Full Text »    PDF »
Role of the Proline-rich Domain of Dynamin-2 and Its Interactions with Src Homology 3 Domains during Endocytosis of the AT1 Angiotensin Receptor.
M. Szaszak, Z. Gaborik, G. Turu, P. S. McPherson, A. J. L. Clark, K. J. Catt, and L. Hunyady (2002)
J. Biol. Chem. 277, 21650-21656
   Abstract »    Full Text »    PDF »
G Protein-Coupled Receptor Allosterism and Complexing.
A. Christopoulos and T. Kenakin (2002)
Pharmacol. Rev. 54, 323-374
   Abstract »    Full Text »    PDF »
{beta}-Adrenergic Signaling in the Heart: Dual Coupling of the {beta}2-Adrenergic Receptor to Gs and Gi Proteins.
R.-P. Xiao (2001)
Sci. STKE 2001, re15
   Abstract »    Full Text »    PDF »
Evolving Concepts in G Protein-Coupled Receptor Endocytosis: The Role in Receptor Desensitization and Signaling.
S. S. G. Ferguson (2001)
Pharmacol. Rev. 53, 1-24
   Abstract »    Full Text »    PDF »
Characterization of ANKRA, a Novel Ankyrin Repeat Protein that Interacts with the Cytoplasmic Domain of Megalin.
K. RADER, R. A. ORLANDO, X. LOU, and M. G. FARQUHAR (2000)
J. Am. Soc. Nephrol. 11, 2167-2178
   Abstract »    Full Text »
Aryloxypropanolamine and Catecholamine Ligand Interactions with the beta 1-Adrenergic Receptor: Evidence for Interaction with Distinct Conformations of beta 1-Adrenergic Receptors.
A. A. Konkar, Z. Zhu, and J. G. Granneman (2000)
J. Pharmacol. Exp. Ther. 294, 923-932
   Abstract »    Full Text »
Catecholamines, Cardiac {beta}-Adrenergic Receptors, and Heart Failure.
R. J. Lefkowitz, H. A. Rockman, and W. J. Koch (2000)
Circulation 101, 1634-1637
   Full Text »    PDF »
Multiple endocytic pathways of G protein-coupled receptors delineated by GIT1 sensitivity.
A. Claing, S. J. Perry, M. Achiriloaie, J. K. L. Walker, J. P. Albanesi, R. J. Lefkowitz, and R. T. Premont (2000)
PNAS 97, 1119-1124
   Abstract »    Full Text »    PDF »
beta 1-Adrenergic Receptor Association with PSD-95. INHIBITION OF RECEPTOR INTERNALIZATION AND FACILITATION OF beta 1-ADRENERGIC RECEPTOR INTERACTION WITH N-METHYL-D-ASPARTATE RECEPTORS.
L. A. Hu, Y. Tang, W. E. Miller, M. Cong, A. G. Lau, R. J. Lefkowitz, and R. A. Hall (2000)
J. Biol. Chem. 275, 38659-38666
   Abstract »    Full Text »    PDF »
G-protein-coupled Receptor Stimulation of the p42/p44 Mitogen-activated Protein Kinase Pathway Is Attenuated by Lipid Phosphate Phosphatases 1, 1a, and 2 in Human Embryonic Kidney 293 Cells.
F. Alderton, P. Darroch, B. Sambi, A. McKie, I. S. Ahmed, N. Pyne, and S. Pyne (2001)
J. Biol. Chem. 276, 13452-13460
   Abstract »    Full Text »    PDF »
Endophilin Regulates JNK Activation through Its Interaction with the Germinal Center Kinase-like Kinase.
A. R. Ramjaun, A. Angers, V. Legendre-Guillemin, X.-K. Tong, and P. S. McPherson (2001)
J. Biol. Chem. 276, 28913-28919
   Abstract »    Full Text »    PDF »
Binding of the beta 2 Adrenergic Receptor to N-Ethylmaleimide-sensitive Factor Regulates Receptor Recycling.
M. Cong, S. J. Perry, L. A. Hu, P. I. Hanson, A. Claing, and R. J. Lefkowitz (2001)
J. Biol. Chem. 276, 45145-45152
   Abstract »    Full Text »    PDF »
Differential Expression of Endophilin 1 and 2 Dimers at Central Nervous System Synapses.
N. Ringstad, Y. Nemoto, and P. De Camilli (2001)
J. Biol. Chem. 276, 40424-40430
   Abstract »    Full Text »    PDF »
G Protein-coupled Receptor Kinase 5 Regulates beta 1-Adrenergic Receptor Association with PSD-95.
L. A. Hu, W. Chen, R. T. Premont, M. Cong, and R. J. Lefkowitz (2002)
J. Biol. Chem. 277, 1607-1613
   Abstract »    Full Text »    PDF »
beta 1-Adrenergic Receptor Association with the Synaptic Scaffolding Protein Membrane-associated Guanylate Kinase Inverted-2 (MAGI-2). DIFFERENTIAL REGULATION OF RECEPTOR INTERNALIZATION BY MAGI-2 AND PSD-95.
J. Xu, M. Paquet, A. G. Lau, J. D. Wood, C. A. Ross, and R. A. Hall (2001)
J. Biol. Chem. 276, 41310-41317
   Abstract »    Full Text »    PDF »

ADVERTISEMENT
Click Me!

ADVERTISEMENT
Click Me!

To Advertise     Find Products

Science Signaling. ISSN 1937-9145 (pre-2008: Science's STKE. ISSN 1525-8882)