Characterization of pancreatic somatostatin binding sites with a 125I-somatostatin 28 analog

Peptides. 1986 Nov-Dec;7(6):953-9. doi: 10.1016/0196-9781(86)90120-8.

Abstract

Somatostatin binding to guinea pig pancreatic acinar cell plasma membranes was characterized with an iodinated stable analog of somatostatin 28 (S28): 125I-[Leu8,DTrp22,Tyr25]S28. The binding was highly dependent on calcium ions. In 0.2 mM free Ca2+ medium, binding at 37 degrees C was saturable, slowly reversible and exhibited a single class of high affinity binding sites (KD = 0.05 +/- 0.01 nM, Bmax = 157 +/- 33 fmol/mg protein). Dissociation of bound radioactivity occurred with biphasic kinetics. Rate of dissociation increased when dissociation was measured at a time before equilibrium binding was reached. In 30 nM free Ca2+ medium, binding affinity and maximal binding capacity were decreased by about 4-fold. Decreasing calcium concentrations increased the amount of rapidly dissociating form of the receptor. Somatostatin 14 antagonist, Des AA1,2[AzaAla4-5,DTrp8, Phe12-13]-somatostatin was active at the membrane level in inhibiting the binding. We conclude that using 125I-[Leu8,DTrp22,Tyr25]S28 as radioligand allows us to characterize a population of specific somatostatin receptors which are not different from those we previously described with the radioligand 125I-[Tyr11]-somatostatin. Somatostatin receptors could exist in two interconvertible forms. Calcium ions are an essential component in the regulation of the conformational change of somatostatin receptors.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Binding, Competitive
  • Calcium / pharmacology
  • Cell Membrane / metabolism
  • Guinea Pigs
  • Iodine Radioisotopes
  • Kinetics
  • Male
  • Pancreas / metabolism*
  • Receptors, Neurotransmitter / metabolism*
  • Receptors, Somatostatin
  • Somatostatin / analogs & derivatives
  • Somatostatin / metabolism*
  • Somatostatin-28

Substances

  • Iodine Radioisotopes
  • Receptors, Neurotransmitter
  • Receptors, Somatostatin
  • Somatostatin
  • Somatostatin-28
  • Calcium