| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH |
|
|
|
|||||||
|
|||||||||
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Papers In Press, published online ahead of print October 22, 2004
Department of Medicine, HHMI/Duke University Medical Center, Durham, NC 27710
Corresponding Author: lefko001{at}receptor-biol.duke.edu
J. Biol. Chem, 10.1074/jbc.M409785200
Submitted on August 25, 2004
Revised on October 19, 2004
Accepted on October 22, 2004
Activation dependent conformational changes in
-arrestin 2
-arrestins are multifunctional adaptor proteins which mediate desensitization, endocytosis and alternate signaling pathways of seven-membrane-spanning receptors (7MSRs). Crystal structures of the basal inactive state of visual arrestin (arrestin 1) and -arrestin 1 (arrestin 2) have been resolved. However, little is known about the conformational changes that occur in -arrestins upon binding to the activated phosphorylated receptor. Here we characterize the conformational changes in -arrestin 2 (arrestin 3) by comparing the limited tryptic proteolysis patterns and Matrix Assisted Laser Desorption/Ionization-Time of Flight mass spectrometry (MALDI-TOF MS) profiles of -arrestin 2 in the presence of a phosphopeptide (V2R-pp) derived from the C-terminus of the vasopressin type II receptor (V2R) or the corresponding nonphosphopeptide (V2R-np). V2R-pp binds to -arrestin 2 specifically, whereas V2R-np does not. Activation of -arrestin 2 upon V2R-pp binding involves the release of its C-terminus, as indicated by exposure of a previously inaccessible cleavage site, one of the polar core residues R394, and rearrangement of its N-terminus, as indicated by the shielding of a previously accessible cleavage site, residue R8. Interestingly, binding of the polyanion heparin also leads to release of the C-terminus of -arrestin 2; however, heparin and V2R-pp have different binding site(s) and/or induce different conformational changes in -arrestin 2. Release of the C-terminus from the rest of -arrestin 2 has functional consequences in that it increases the accessibility of a previously defined clathrin binding site(residues 371-379) thereby enhancing clathrin binding to -arrestin 2 by ten-fold. Thus, the V2R-pp can activate -arrestin 2 in vitro, most likely mimicking the effects of an activated phosphorylated 7MSR. These results provide the first direct evidence of conformational changes associated with the transition of -arrestin 2 from its basal inactive conformation to its biologically active conformation and establish a system in which receptor--arrestin interactions can be modeled in vitro.
CiteULike 
Complore 
Connotea 
Del.icio.us 
Digg 
Reddit 
Technorati What's this?
This article has been cited by other articles:
|
|
 |
|
  S. K. Shenoy, K. Xiao, V. Venkataramanan, P. M. Snyder, N. J. Freedman, and A. M. Weissman Nedd4 Mediates Agonist-dependent Ubiquitination, Lysosomal Targeting, and Degradation of the {beta}2-Adrenergic Receptor J. Biol. Chem., August 8, 2008; 283(32): 22166 - 22176. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. D. Nelson, J. J. Kovacs, K. N. Nobles, E. J. Whalen, and R. J. Lefkowitz {beta}-Arrestin Scaffolding of Phosphatidylinositol 4-Phosphate 5-Kinase I{alpha} Promotes Agonist-stimulated Sequestration of the {beta}2-Adrenergic Receptor J. Biol. Chem., July 25, 2008; 283(30): 21093 - 21101. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. K. Shukla, J. D. Violin, E. J. Whalen, D. Gesty-Palmer, S. K. Shenoy, and R. J. Lefkowitz Distinct conformational changes in {beta}-arrestin report biased agonism at seven-transmembrane receptors PNAS, July 22, 2008; 105(29): 9988 - 9993. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 L. Barki-Harrington and H. A. Rockman {beta}-Arrestins: Multifunctional Cellular Mediators Physiology, February 1, 2008; 23(1): 17 - 22. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 T. Kenakin Functional Selectivity through Protean and Biased Agonism: Who Steers the Ship? Mol. Pharmacol., December 1, 2007; 72(6): 1393 - 1401. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. Qiu, H. H. Loh, and P.-Y. Law Phosphorylation of the {delta}-Opioid Receptor Regulates Its beta-Arrestins Selectivity and Subsequent Receptor Internalization and Adenylyl Cyclase Desensitization J. Biol. Chem., August 3, 2007; 282(31): 22315 - 22323. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. N. Nobles, Z. Guan, K. Xiao, T. G. Oas, and R. J. Lefkowitz The Active Conformation of beta-Arrestin1: DIRECT EVIDENCE FOR THE PHOSPHATE SENSOR IN THE N-DOMAIN AND CONFORMATIONAL DIFFERENCES IN THE ACTIVE STATES OF beta-ARRESTINS1 AND -2 J. Biol. Chem., July 20, 2007; 282(29): 21370 - 21381. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. M. Hanson, D. J. Francis, S. A. Vishnivetskiy, E. A. Kolobova, W. L. Hubbell, C. S. Klug, and V. V. Gurevich Differential interaction of spin-labeled arrestin with inactive and active phosphorhodopsin PNAS, March 28, 2006; 103(13): 4900 - 4905. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. A. Willoughby and M. K. Collins Dynamic Interaction between the Dual Specificity Phosphatase MKP7 and the JNK3 Scaffold Protein {beta}-Arrestin 2 J. Biol. Chem., July 8, 2005; 280(27): 25651 - 25658. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
X.-R. Ren, E. Reiter, S. Ahn, J. Kim, W. Chen, and R. J. Lefkowitz Different G protein-coupled receptor kinases govern G protein and {beta}-arrestin-mediated signaling of V2 vasopressin receptor PNAS, February 1, 2005; 102(5): 1448 - 1453. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH |
| All ASBMB Journals | Molecular and Cellular Proteomics |
| Journal of Lipid Research | ASBMB Today |
