Solubilization, characterization and partial purification of (3H)mepyramine-binding protein, a possible histamine H1 receptor, from rat liver membrane.

DOI PubMed Open Access
  • FUKUI Hiroyuki
    Department of Pharmacology II, Osaka University School of Medicine
  • WANG Nai Ping
    Department of Pharmacology II, Osaka University School of Medicine
  • WATANABE Takehiko
    Department of Pharmacology I, Tohoku University School of Medicine
  • WADA Hiroshi
    Department of Pharmacology II, Osaka University School of Medicine

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Description

[3H] Mepyramine binding protein, a possible subtype of histamine H, receptors, was solubilized from rat liver membrane with 3-[(3-cholamidopropyl)-dimethylammonio] -1 -propanesulfonate (CHAPS) and Tween 60 as detergents and glycerol as an enhancer of solubilization. The optimal concentration of CHAPS was 10 mM and that of glycerol was 20% or more (v/v). The molecular weight of the [3H]mepyramine binding protein-detergent complex was determined to be 670K by Sepharose CL-4B gel filtration and 800K by sucrose density gradient sedimentation. By target size analysis, the molecular weights of both the membranebound and solubilized [3H]mepyramine binding protein were determined to be 162K. These values are similar to those of other well-characterized H, -receptor proteins, though slightly different. Simultaneous computerized analysis of the data obtained by [3H]mepyramine binding to the solubilized [3H]mepyramine binding protein indicated the presence of a single binding site with a KD value of 19.0±5.6 nM and a binding capacity (Bmax) of 6.6±2.1 pmole/mg protein. The Ki value of cold mepyramine for [3H]mepyramine binding to the solubilized receptor was 20±4 nM, whereas those of diphenhydramine, d-chlorpheniramine and triprolidine were all 2.9±0.8 μM, or about 150 times that of mepyramine. These data on the molecular and binding characteristics of the solubilized protein reported here suggest that there is a subtype of histamine H1 receptor in rat liver membrane. The solubilized preparation retained 90% and 75% of its [3H]mepyramine binding activity after storage at -80°C and 4°C, respectively, for 20 days. The solubilized [3H]mepyramine binding protein was purified 30-fold by Sepharose CL-4B gel filtration, Bio Gel HTP hydroxylapatite, Octyl Sepharose 4B and hydroxylapatite HPLC column chromatographies.

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Details 詳細情報について

  • CRID
    1390001204289103616
  • NII Article ID
    130000832101
  • DOI
    10.1254/jjp.46.127
  • COI
    1:CAS:528:DyaL1cXhtVKqtL8%3D
  • ISSN
    13473506
    00215198
  • PubMed
    3379823
  • Text Lang
    en
  • Article Type
    journal article
  • Data Source
    • JaLC
    • Crossref
    • PubMed
    • CiNii Articles
    • OpenAIRE
  • Abstract License Flag
    Disallowed

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