Isolation of Substance P Binding Protein from Rat Brain.

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概要

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• Substance P (SP) binding protein of rat brain was solubilized by digitonin. The solubilized proteins were then purified by sequential gel filtration, concanavalin A lectin Sepharose, and SP-affinity chromatography. The calculated molecular weight of this purified SP binding protein was 76-74 kDa on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The rabbits were immunized with the purified protein and resulting polyclonal anti-sera were tested. The immune serum significantly inhibited [<SUP>3</SUP>H]SP binding to the 3-[(3-cholamidopropyl)dimethylammonio]-1-propane sulfonate solubilized membrane fractions from rat brain, whereas pre-bleed antiserum failed to inhibit the binding. This polyclonal antibody also inhibited the activity of <SUP>45</SUP>Ca influx into astroglioma cells stimulated by SP, but does not inhibit that stimulated by histamine. Furthermore, this polyclonal antibody recognized the 76-74 kDa band as assessed by Western blotting. These data strongly suggest that this polyclonal antibody could recognize a part of the natural SP receptor site.

• 公益社団法人 日本薬理学会の論文

著者

• Nishio Hiroaki

  Department Of Molecular Pharmacology Faculty Of Pharmacy And Pharmaceutical Sciences Fukuyama Univer

• Hirai Yuko

  Department Of Cerebrovascular Disease National Kyushu Medical Center

• Akiyama Mitoshi

  Department Of Clinical Radiology Faculty Health Sciences Hiroshima International University

• AKIZAWA Toshifumi

  Faculty of Pharmaceutical Sciences, Setsunan University

• Nakata Yoshihiro

  Department Of Biophysics Gunma University

• Akizawa Toshifumi

  Faculty Of Pharmaceutical Sciences Setsunan University

• TAKAMATSU Hiroyuki

  Department of Cellular Transplantation Biology, Kanazawa University Graduate School of Medicine

• Hirai Yuko

  Department of Radiobiology, Radiation Effects Research Foundation

• Kuroyanagi Naoyoshi

  Department of Pharmacology, Institute of Pharmaceutical Sciences, Hiroshima University School of Medicine

• Segawa Tomio

  Tsuruga Women''s Junior College

• Segawa Tomio

  Tsuruga Women''s Junior College

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