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. 1988 Apr;85(8):2743–2747. doi: 10.1073/pnas.85.8.2743

Characterization of histamine H1-receptor binding peptides in guinea pig brain using [125I]iodoazidophenpyramine, an irreversible specific photoaffinity probe.

M Ruat 1, M Körner 1, M Garbarg 1, C Gros 1, J C Schwartz 1, W Tertiuk 1, C R Ganellin 1
PMCID: PMC280075  PMID: 3357888

Abstract

Aminophenpyramine--i.e., N-(5-[2-(4-aminophenyl)ethanamidopentyl])-N'-(4-methoxybenzyl)-N-m ethyl-N'- (2-pyridinyl)-1,2-ethanediamine, a derivative of mepyramine (pyrilamine), a typical antagonist of histamine at its H1 receptor--was synthesized and converted into [125I)iodoazidophenpyramine, a potential photoaffinity probe for the H1 receptor. In the dark, reversible binding of this probe to cerebellar membranes occurred with a Kd of 1.2 x 10(-11) M and a Bmax of 240 fmol/mg of protein and was inhibited by various H1-receptor antagonists with the expected potencies. These features establish the compound as one of the most potent H1-receptor antagonists known so far. Upon UV irradiation, 5% of the bound radioactivity was covalently incorporated into cerebellar membrane polypeptides as shown by standard NaDodSO4/PAGE. Two bands of 47 and 56 kDa were consistently labeled, labeling being prevented by various H1-receptor antagonists with the expected potencies and stereoselectivity. In the presence of protease inhibitors, labeling of the 56-kDa peptide increased at the expense of the 47-kDa peptide, suggesting that the latter was produced by hydrolysis of the former under the action of membrane proteases. In the absence of 2-mercaptoethanol, a band of 350-400 kDa appeared, apparently at the expense of the lighter bands, suggesting that the latter might be linked by one or more disulfide bridges to a higher molecular mass complex. We propose that at least part of the ligand binding domain of the histamine H1 receptor resides within a subunit of apparent molecular mass 56,000.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Chang R. S., Synder S. H. Histamine H1-receptor binding sites in guinea pig brain membranes: regulation of agonist interactions by guanine nucleotides and cations. J Neurochem. 1980 Apr;34(4):916–922. doi: 10.1111/j.1471-4159.1980.tb09666.x. [DOI] [PubMed] [Google Scholar]
  2. Donaldson J., Hill S. J. 1,4-Dithiothreitol-induced alteration in histamine H1-agonist binding in guinea-pig cerebellum and cerebral cortex. Eur J Pharmacol. 1986 Sep 23;129(1-2):25–31. doi: 10.1016/0014-2999(86)90332-8. [DOI] [PubMed] [Google Scholar]
  3. Hill S. J., Young J. M., Marrian D. H. Specific binding of 3H-mepyramine to histamine H1 receptors in intestinal smooth muscle. Nature. 1977 Nov 24;270(5635):361–363. doi: 10.1038/270361a0. [DOI] [PubMed] [Google Scholar]
  4. Korner M., Bouthenet M. L., Ganellin C. R., Garbarg M., Gros C., Ife R. J., Sales N., Schwartz J. C. [125I]Iodobolpyramine, a highly sensitive probe for histamine H1-receptors in guinea-pig brain. Eur J Pharmacol. 1986 Jan 21;120(2):151–160. doi: 10.1016/0014-2999(86)90535-2. [DOI] [PubMed] [Google Scholar]
  5. Kuno T., Kubo N., Tanaka C. Molecular size of histamine H-1 receptor determined by target size analysis. Biochem Biophys Res Commun. 1985 Jun 28;129(3):639–644. doi: 10.1016/0006-291x(85)91939-4. [DOI] [PubMed] [Google Scholar]
  6. Porzio M. A., Pearson A. M. Improved resolution of myofibrillar proteins with sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Biochim Biophys Acta. 1977 Jan 25;490(1):27–34. doi: 10.1016/0005-2795(77)90102-7. [DOI] [PubMed] [Google Scholar]
  7. Toll L., Snyder S. H. Solubilization and Characterization of Histamine H1 receptors in brain. J Biol Chem. 1982 Nov 25;257(22):13593–13601. [PubMed] [Google Scholar]
  8. Tran V. T., Chang R. S., Snyder S. H. Histamine H1 receptors identified in mammalian brain membranes with [3H]mepyramine. Proc Natl Acad Sci U S A. 1978 Dec;75(12):6290–6294. doi: 10.1073/pnas.75.12.6290. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Wang N. P., Fukui H., Matsuoka H., Wada H. Determination of the molecular size of the hepatic H1-receptor by target size analysis. Biochem Biophys Res Commun. 1986 Jun 13;137(2):593–598. doi: 10.1016/0006-291x(86)91119-8. [DOI] [PubMed] [Google Scholar]
  10. Waud D. R., Parker R. B. Pharmacological estimation of drug-receptor dissociation constants. Statistical evaluation. II. Competitive antagonists. J Pharmacol Exp Ther. 1971 Apr;177(1):13–24. [PubMed] [Google Scholar]
  11. Wouters W., Van Dun J., Leysen J. E., Laduron P. M. Differential photoaffinity labelling of serotonin-S2 receptors and histamine-H1 receptors using 7-azidoketanserin. FEBS Lett. 1985 Mar 25;182(2):291–296. doi: 10.1016/0014-5793(85)80318-5. [DOI] [PubMed] [Google Scholar]
  12. Wouters W., Van Dun J., Leysen J. E., Laduron P. M. Photoaffinity probes for serotonin and histamine receptors. Synthesis and characterization of two azide analogues of ketanserin. J Biol Chem. 1985 Jul 15;260(14):8423–8429. [PubMed] [Google Scholar]
  13. Yeramian E., Garbarg M., Schwartz J. C. N-ethylmaleimide-induced changes in agonist affinity for histamine H1-receptors in the guinea pig brain. Mol Pharmacol. 1985 Aug;28(2):155–162. [PubMed] [Google Scholar]

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