Elucidation of inositol hexaphosphate and heparin interaction sites and conformational changes in arrestin-1 by solution nuclear magnetic resonance.
Arrestins specifically bind activated and phosphorylated G protein-coupled receptors and orchestrate both receptor trafficking and channel signaling through G protein-independent pathways via direct interactions with numerous nonreceptor partners. Here we report the first successful use of solution NMR in mapping the binding sites in arrestin-1 (visual arrestin) for two polyanionic compounds that mimic phosphorylated light-activated rhodopsin: inositol hexaphosphate (IP6) and heparin. This yielded an identification of residues involved in the binding with these ligands that was more complete than what has previously been feasible. IP6 and heparin appear to bind to the same site on arrestin-1, centered on a positively charged region in the N-domain. We present the first direct evidence that both IP6 and heparin induced a complete release of the arrestin C-tail. These observations provide novel insight into the nature of the transition of arrestin from the basal to active state and demonstrate the potential of NMR-based methods in the study of protein-protein interactions involving members of the arrestin family.
Department of Biochemistry, Vanderbilt University School ofMedicine, Nashville, Tennessee 37232, United States.
Tiandi
T
Zhuang
Sergey A
SA
Vishnivetskiy
Vsevolod V
VV
Gurevich
Charles R
CR
Sanders
D000595:Amino Acid Sequence; D000818:Animals; D019390:Arrestins / Q000737:chemistry / Q000378:metabolism*; D001665:Binding Sites / Q000502:physiology; D002417:Cattle; D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000235:genetics; D006493:Heparin / Q000737:chemistry / Q000378:metabolism*; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D008969:Molecular Sequence Data; D010833:Phytic Acid / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding / Q000502:physiology; D011487:Protein Conformation; D011817:Rabbits; D012996:Solutions
D019390:Arrestins; D012996:Solutions; D010833:Phytic Acid; D006493:Heparin