Key differences in molecular complexes of the cholecystokinin receptor with structurally related peptide agonist, partial agonist, and antagonist

Abstract

The molecular basis of docking of receptor ligands having differences in biological activity and their subsequent effects on receptor conformation represent areas of great interest. In this work, we focus on the sulfated tyrosyl residue in position 27 of cholecystokinin (CCK) and its spatial approximation with the type A CCK receptor residue Arg(197) that has been predicted from mutagenesis experiments. We have examined the requirement for sulfation of this residue in a series of structurally related peptide agonists, partial agonists, and antagonists using assays of receptor binding and biological activity. Whereas sulfation of CCK position 27 was critical for affinity and potency of a full agonist, it had progressively less effect as the biological activity of the ligand was reduced. It had an intermediate effect on the partial agonist and no effect on the antagonist. In addition, photoaffinity labeling was used to determine the spatial approximations between the receptor and residue 27 of the agonist and antagonist in this series. Direct photoaffinity labeling with a full agonist probe confirmed the spatial approximation of ligand residue 27 and receptor residue Arg(197) in the active complex. Of note, the analogous antagonist probe labeled a distinct region within the receptor amino terminus, confirming a key structural difference in active and inactive complexes.