A mnemonical or negative-co-operativity model for the activation of adenylate cyclase by a common G-protein-coupled calcitonin-gene-related neuropeptide (CGRP)/amylin receptor

Biochem J. 1993 Jul 1;293 ( Pt 1)(Pt 1):229-36. doi: 10.1042/bj2930229.

Abstract

Both amylin and calcitonin-gene-related neuropeptide (CGRP) activated adenylate cyclase activity in hepatocyte membranes around 5-fold in a dose-dependent fashion, with EC50 values of 120 +/- 14 and 0.3 +/- 0.14 nM respectively. Whereas amylin exhibited normal activation kinetics (Hill coefficient, h approximately 1), CGRP showed kinetics indicative of either multiple sites/receptor species having different affinities for this ligand or a single receptor species exhibiting apparent negative co-operativity (h approximately 0.21). The CGRP antagonist CGRP-(8-37)-peptide inhibited adenylate cyclase stimulated by EC50 concentrations of either amylin or CGRP. Inhibition by CGRP-(8-37) was selective in that markedly lower concentrations were required to block the action of amylin (IC50 = 3 +/- 1 nM) compared with that of CGRP itself (IC50 = 120 +/- 11 nM). Dose-effect data for inhibition of CGRP action by CGRP-(8-37) showed normal saturation kinetics (h approximately 1), whereas CGRP-(8-37) inhibited amylin-stimulated adenylate cyclase activity in a fashion which was indicative of either multiple sites or apparent negative co-operativity (h approximately 0.24). Observed changes in the kinetics of inhibition by CGRP-(8-37) of CGRP, but not amylin-stimulated adenylate cyclase, at concentrations of agonists below their EC50 values militated against a model of two distinct populations of non-interacting receptors each able to bind both amylin and CGRP. A kinetic model is proposed whereby a single receptor, capable of being activated by both CGRP and amylin, obeys either a mnemonical kinetic mechanism or one of negative co-operativity with respect to CGRP but not to amylin. The relative merits of these two models are discussed together with a proposal suggesting that the activation of adenylate cyclase by various G-protein-linked receptors may be described by a mnemonical model mechanism.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenylyl Cyclases / metabolism*
  • Amyloid / pharmacology*
  • Animals
  • Calcitonin Gene-Related Peptide / pharmacology*
  • Cyclic AMP / metabolism
  • Enzyme Activation
  • GTP-Binding Proteins / metabolism*
  • Humans
  • Islet Amyloid Polypeptide
  • Kinetics
  • Liver / cytology
  • Liver / drug effects
  • Liver / enzymology*
  • Male
  • Models, Biological
  • Rats
  • Rats, Sprague-Dawley
  • Receptors, Calcitonin
  • Receptors, Cell Surface / metabolism
  • Receptors, Islet Amyloid Polypeptide
  • Receptors, Peptide*

Substances

  • Amyloid
  • Islet Amyloid Polypeptide
  • Receptors, Calcitonin
  • Receptors, Cell Surface
  • Receptors, Islet Amyloid Polypeptide
  • Receptors, Peptide
  • Cyclic AMP
  • GTP-Binding Proteins
  • Adenylyl Cyclases
  • Calcitonin Gene-Related Peptide