Lactam constraints provide insights into the receptor-bound conformation of secretin and stabilize a receptor antagonist

Maoqing Dong, Jerez A. Te, Xiequn Xu, Jinhui Wang, Delia I. Pinon, Laura Storjohann, Andrew J. Bordner, Laurence J. Miller

Research output: Contribution to journalArticle

13 Scopus citations

Abstract

The natural ligands for family B G protein-coupled receptors are moderate-length linear peptides having diffuse pharmacophores. The amino-terminal regions of these ligands are critical for biological activity, with their amino-terminal truncation leading to production of orthosteric antagonists. The carboxyl-terminal regions of these peptides are thought to occupy a ligand-binding cleft within the disulfide-bonded amino-terminal domains of these receptors, with the peptides in amphipathic helical conformations. In this work, we have characterized the binding and activity of a series of 11 truncated and lactam-constrained secretin(5-27) analogues at the prototypic member of this family, the secretin receptor. One peptide in this series with lactam connecting residues 16 and 20 [c[E 16,K 20][Y 10]sec(5-27)] improved the binding affinity of its unconstrained parental peptide 22-fold while retaining the absence of endogenous biological activity and competitive antagonist characteristics. Homology modeling with molecular mechanics and molecular dynamics simulations established that this constrained peptide occupies the ligand-binding cleft in an orientation similar to that of natural full-length secretin and provided insights into why this peptide was more effective than other truncated conformationally constrained peptides in the series. This lactam bridge is believed to stabilize an extended α-helical conformation of this peptide while in solution and not to interfere with critical residue-residue approximations while docked to the receptor.

Original languageEnglish (US)
Pages (from-to)8181-8192
Number of pages12
JournalBiochemistry
Volume50
Issue number38
DOIs
StatePublished - Sep 27 2011

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ASJC Scopus subject areas

  • Biochemistry

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