High-affinity [3H]pirenzepine binding to putative M1 muscarinic sites in the neuroblastoma × glioma hybrid cell line (NG 108-15)

Kazufumi Akiyama, Mark Watson, William R Roeske, Henry I. Yamamura

Research output: Contribution to journalArticle

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Abstract

The specific binding of both the non-classical antagonist [3H] pirenzepine ([3H]PZ) and the classical antagonist [3H](-)quinuclidinyl benzilate ([3H](-)QNB) was determined in parallel assays of the mouse neuroblastoma × rat glioma hybrid cell line (NG 108-15). Saturation isotherms yielded a Kd = 4.0 nM and Bmax = 27.8 fmoles/mg protein for [3H]PZ and a Kd = 17.2 pM and Bmax = 53.2 fmoles/mg protein for [3H](-)QNB. The inhibition data of pirenzepine vs [3H](-)QNB was best fit to a 2-site binding model revealing both a high affinity pirenzepine site (72%, KH = 10.3 nM) and a low affinity site (28%, KL = 97.5 nM). [3H]PZ competition studies demonstrated stereospecificity, steep inhibition curves for muscarinic antagonists (Hill coefficients close to 1), and a shallow inhibition curve for a muscarinic agonist. These results indicate that muscarinic receptors on NG 108-15 cells may be subclassified (M1/M2) on the basis of the discriminative capability of [3H]PZ.

Original languageEnglish (US)
Pages (from-to)289-297
Number of pages9
JournalBiochemical and Biophysical Research Communications
Volume119
Issue number1
DOIs
Publication statusPublished - Feb 29 1984

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ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

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