Identification and characterization of calcitonin gene-related peptide receptors in porcine renal medullary membranes

Endocrinology. 1991 Aug;129(2):965-9. doi: 10.1210/endo-129-2-965.

Abstract

Membranes prepared from the medullary region of the porcine kidney displayed high affinity, high density (Kd, 0.12 nM; binding capacity, 127 fmol/mg protein) receptors for calcitonin gene-related peptide (CGRP). Human CGRP (hCGRP), rat CGRP (rCGRP), and the hCGRP analog [hCGRP-(8-37)] competed for the binding of [125I]hCGRP, whereas salmon calcitonin (sCT) and CGRP-(22-37) were very weak in displacing [125I]hCGRP binding. In accordance with these binding data, CGRP stimulated adenylate cyclase activity in these membrane preparations in a concentration-dependent manner, with an EC50 similar to that of the Kd for binding. In the same preparations, sCT was ineffective in stimulating adenylate cyclase activity, suggesting that porcine kidney medullary membranes possess receptors specific for CGRP. Further hCGRP-(8-37), a CGRP antagonist, inhibited CGRP-stimulated adenylate cyclase activity in a competitive manner. Covalent cross-linking of [125I]hCGRP to these membranes resulted in the specific labeling of one major band at approximately 30,000 mol wt and two minor bands at about 58,000 and 78,000 mol wt. The presence of CGRP receptors and their coupling to adenylate cyclase suggest a role for CGRP in kidney function, such as local regulation of the microcirculation, electrolyte transport, or water homeostasis in the porcine kidney.

MeSH terms

  • Adenylyl Cyclases / metabolism*
  • Animals
  • Binding, Competitive
  • Calcitonin Gene-Related Peptide / metabolism
  • Calcitonin Gene-Related Peptide / pharmacology
  • Cell Membrane / metabolism
  • Cross-Linking Reagents
  • Humans
  • Kidney Medulla / metabolism*
  • Molecular Weight
  • Rats
  • Receptors, Calcitonin
  • Receptors, Cell Surface / analysis
  • Receptors, Cell Surface / chemistry
  • Receptors, Cell Surface / metabolism*
  • Second Messenger Systems
  • Swine

Substances

  • Cross-Linking Reagents
  • Receptors, Calcitonin
  • Receptors, Cell Surface
  • Adenylyl Cyclases
  • Calcitonin Gene-Related Peptide