NMR analysis of rhodopsin-transducin interactions.
Heterotrimeric G-protein activation by an agonist-stimulated G-protein coupled receptor (R*) requires the propagation of structural signals from the receptor interacting surfaces to the guanine nucleotide-binding pocket. Employing high-resolution NMR methods, we are probing heterotrimer-associated and rhodopsin-stimulated changes in an isotope-labeled G-protein alpha-subunit (G(alpha)). A key aspect of the work involves the trapping and interrogation of discrete R*-bound conformations of G(alpha). Our results demonstrate that functionally important changes in G(alpha) structure and dynamics can be detected and characterized by NMR, enabling the generation of robust models for the global and local structural changes accompanying signal transfer from R* to the G-protein.
Ridge
K D
KD
Center for Membrane Biology, Department of Biochemistry and Molecular Biology, University of Texas Health Science Center, Houston, TX 77030, USA. Kevin.D.Ridge@uth.tmc.edu
Marino
J P
JP
Ngo
T
T
Ramon
E
E
Brabazon
D M
DM
Abdulaev
N G
NG
D000818:Animals; D001665:Binding Sites; D002417:Cattle; D019281:Dimerization; D044385:GTP-Binding Protein alpha Subunits / Q000378:metabolism; D007553:Isotope Labeling; D009682:Magnetic Resonance Spectroscopy; D008968:Molecular Conformation; D012243:Rhodopsin / Q000378:metabolism*; D012374:Rod Cell Outer Segment / Q000378:metabolism*; D015293:Transducin / Q000378:metabolism*; D014785:Vision, Ocular / Q000502:physiology
D044385:GTP-Binding Protein alpha Subunits; D012243:Rhodopsin; D015293:Transducin