Direct analysis of a GPCR-agonist interaction by surface plasmon resonance.
Despite their clinical importance, detailed analysis of ligand binding at G-protein coupled receptors (GPCRs) has proved difficult. Here we successfully measure the binding of a GPCR, neurotensin receptor-1 (NTS-1), to its ligand, neurotensin (NT), using surface plasmon resonance (SPR). Specific responses were observed between NT and purified, detergent-solublised, recombinant NTS-1, using a novel configuration where the biotinylated NT ligand was immobilised on the biosensor surface. This SPR approach shows promise as a generic approach for the study of ligand interactions with other suitable GPCRs.
Harding
Peter J
PJ
Biomembrane Structure Unit, Department of Biochemistry, University of Oxford, South Parks Road, Oxford, UK.
Hadingham
Timothy C
TC
McDonnell
James M
JM
Watts
Anthony
A
D000818:Animals; D007700:Kinetics; D008024:Ligands; D009496:Neurotensin / Q000737:chemistry*; D051381:Rats; D018028:Receptors, Neurotensin / Q000819:agonists* / Q000737:chemistry*; D020349:Surface Plasmon Resonance
D008024:Ligands; D018028:Receptors, Neurotensin; C114036:neurotensin type 1 receptor; D009496:Neurotensin