title: Evidence that the Rhodopsin Kinase (GRK1) N-Terminus and the Transducin Gα C-Terminus Interact with the Same "Hydrophobic Patch" on Rhodopsin TM5.
abstract: Phosphorylation of G protein-coupled receptors (GPCRs) terminates their ability to couple with and activate G proteins by increasing their affinity for arrestins. Unfortunately, detailed information regarding how GPCRs interact with the kinases responsible for their phosphorylation is still limited. Here, we purified fully functional GPCR kinase 1 (GRK1) using a rapid method and used it to gain insights into how this important kinase interacts with the GPCR rhodopsin. Specifically, we find that GRK1 uses the same site on rhodopsin as the transducin (Gt) Gtα C-terminal tail and the arrestin "finger loop", a cleft formed in the cytoplasmic face of the receptor upon activation. Our studies also show GRK1 requires two conserved residues located in this cleft (L226 and V230) that have been shown to be required for Gt activation due to their direct interactions with hydrophobic residues on the Gα C-terminal tail. Our data and modeling studies are consistent with the idea that all three proteins (Gt, GRK1, and visual arrestin) bind, at least in part, in the same site on rhodopsin and interact with the receptor through a similar hydrophobic contact-driven mechanism.
authors.affiliation: Department of Biochemistry and Molecular Biology, Oregon Health and Science University , Portland, Oregon 97239-3098, United States.
authors.forename: Amber M
authors.initials: AM
authors.lastname: Jones Brunette
authors.affiliation: Department of Biochemistry and Molecular Biology, Oregon Health and Science University , Portland, Oregon 97239-3098, United States.
authors.forename: Abhinav
authors.initials: A
authors.lastname: Sinha
authors.affiliation: Department of Biochemistry and Molecular Biology, Oregon Health and Science University , Portland, Oregon 97239-3098, United States.
authors.forename: Larry
authors.initials: L
authors.lastname: David
authors.affiliation: Department of Biochemistry and Molecular Biology, Oregon Health and Science University , Portland, Oregon 97239-3098, United States.
authors.forename: David L
authors.initials: DL
authors.lastname: Farrens
mesh_terms: D019390:Arrestins / Q000737:chemistry / Q000378:metabolism*; D051606:G-Protein-Coupled Receptor Kinase 1 / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D010766:Phosphorylation; D011485:Protein Binding; D011487:Protein Conformation; D043562:Receptors, G-Protein-Coupled / Q000737:chemistry / Q000378:metabolism*; D012243:Rhodopsin / Q000737:chemistry / Q000378:metabolism*; D015398:Signal Transduction; D015293:Transducin / Q000737:chemistry / Q000378:metabolism*
chemical_list: D019390:Arrestins; D043562:Receptors, G-Protein-Coupled; D012243:Rhodopsin; D051606:G-Protein-Coupled Receptor Kinase 1; C496764:GRK1 protein, human; D015293:Transducin