Demonstration of specific binding sites for pituitary adenylate cyclase activating polypeptide (PACAP) in rat astrocytes

Ichiro Tatsuno, Paul E. Gottschall, K. Köves, Akira Arimura

Research output: Contribution to journalArticle

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Abstract

The high and low affinity binding sites for PACAP were identified in rat astrocytes using [125I]PACAP27 as the labeled ligand. Scatchard analysis of displacement of the bound tracer by unlabeled PACAP27 indicated the existence of two classes of binding sites, with the dissociation constant (Kd) = 1.22±0.4 nM, the binding maximal capacity (Bmax) = 821±218 fmols/mg protein for the high affinity binding site, and Kd=0.59±0.06 μM, Bmax=563±12 pmols/mg protein for the low affinity binding site, respectively. The specificity of [125I]PACAP27 binding was tested using PACAP38 and peptides structually related to PACAP, such as VIP, GHRF, PHI, secretin and glucagon. PACAP38 completely displaced the binding of [125I]PACAP27 and Scatchard analysis also indicated the presence of two classes of binding sites with similar Kd and Bmax to those for PACAP27. VIP and GHRF competed with [125I]PACAP27, but to a much lesser extent than unlabeled PACAP27 in binding. Other peptides tested did not displace the binding of [125I]PACAP27 at 10-6 M.

Original languageEnglish
Pages (from-to)1027-1033
Number of pages7
JournalBiochemical and Biophysical Research Communications
Volume168
Issue number3
DOIs
Publication statusPublished - May 16 1990

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Pituitary Adenylate Cyclase-Activating Polypeptide
Astrocytes
Rats
Demonstrations
Binding Sites
Peptides
Secretin
Glucagon
Proteins

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Demonstration of specific binding sites for pituitary adenylate cyclase activating polypeptide (PACAP) in rat astrocytes. / Tatsuno, Ichiro; Gottschall, Paul E.; Köves, K.; Arimura, Akira.

In: Biochemical and Biophysical Research Communications, Vol. 168, No. 3, 16.05.1990, p. 1027-1033.

Research output: Contribution to journalArticle

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