Characterization of G-Protein Coupled Receptor Kinase Interaction with the Neurokinin-1 Receptor Using Bioluminescence Resonance Energy Transfer

• DOI: 10.1124/mol.107.038877
• Publication Date: 2007-11-07T01:34:49Z
• Authors: Rasmus Jorgensen; Nicholas D. Holliday; Jakob L. Hansen; Milka Vrecl; Anders Heding; Thue W. Schwartz; Christian E. Elling
• Keywords: GABBR1; Interleukin-13 receptor

To analyze the interaction between neurokinin-1 (NK-1) receptor and G-protein coupled kinases (GRKs), we performed bioluminescence resonance energy transfer² (BRET²) measurements family A NK-1 GRK2 GRK5 as well their respective kinase-inactive mutants. We observed agonist induced of both with activated receptor. In saturation experiments, to interact in a monophasic manner while interacted biphasic low affinity phase corresponding for GRK5. Agonist was dependent on intact kinase-activity, whereas high independent kinase activity. were surprised find that BRET² experiments indicated before activation, full-length receptor, but not functional C-terminal tail-truncated is preassociated relatively low-affinity state. demonstrate can compete does suggest GRK5, rather than GRK2, key player competitive regulation GRK subtype specific