1990 Fiscal Year Annual Research Report

Pー糖蛋白質の構造と機能の研究

Research Project

Project/Area Number	02807028
Research Institution	Kagoshima University
Principal Investigator	秋山伸一鹿児島大学, 医学部, 教授 (60117413)
Co-Investigator(Kenkyū-buntansha)	古川龍彦鹿児島大学, 医学部, 助手 (40219100) 澄澤知之鹿児島大学, 医学部, 助手 (90206582)
Keywords	Pー糖蛋白質 / 多剤耐性 / 薬剤結合部位 / 副腎
Research Abstract	1.ウシ副腎のPー糖蛋白質の機能の解析ウシ副腎のPー糖蛋白質の薬剤結合部位について解析するため、ウシ副腎から膜画分を分離し、[ ^3H]アジドピンでフォトアフィニティ-ラベルを行った。130KDaの分子がフォトラベルされ、この分子はPー糖給白質にする単クロ-ン抗体、Cー219で免疫沈降した。このことはウシ副腎のPー糖蛋白質が、アジドピンとの結合部位を有し、その分子量はヒト癌KB細胞のPー糖蛋白質より10KDa以上小さいことを示している。[ ^3H]アジドピンによるフォトラベルは、抗癌剤ビンブラスチンや耐性克服薬剤、レセルピン、ベラパミ-ルで阻害された。ウシ副腎Pー糖蛋白質は、ヒト癌KB細胞のPー糖蛋白質の薬剤結合部位とよく似た薬剤結合部位を持つことが判明した。 2.ウシ副腎中より、Pー糖蛋白質の輸送基質となる物質の抽出について。ウシ副腎をホモジェナイズし、100%メタノ-ルで抽出した。これをヘキサンで分配し、ヘキサン層を除去したあと、酢酸エチルで分配し酢酸エチル層を濃縮乾固した。クロロホルムに溶解し、シリカゲルカラムに注入して1%酢酸、2%メタノ-ル、97%クロロホルムで溶出し、多剤耐性細胞への抗癌剤の蓄積を高める活性を各画分で測定した。活性部分を薄層クロマトグラフィ-で展開し、活性部分を採取して、順相HPLC、逆相HPLCにて精製を試みた。最後のODSー2カラムで、16分附近に活性のあるピ-クが認められたので、この物質の精製を進め、構造決定を行う予定である。

Research Products
(13 results)

All Other

All Publications (13 results)

[Publications] Yasuo Kuwazuru: "Expression of the multidrug transporter,Pーglycoprotein,in chronic myelogenous leukemia cells in blast crisis." British Journal of Haematology. 74. 24-29 (1990)
[Publications] Akihiko Yoshimura: "Novel screening method for agents that overcome classical multidrug resistance in a human cell line." Cancer Letters. 50. 45-51 (1990)
[Publications] Akihiko Yoshimura: "Purification and tissue distribution of human thymidine phosphorylase;high expression in lymphocytes,reticulocytes and tumors." Biochimica Biophysica Acta. 1034. 107-113 (1990)
[Publications] Shunーichi Ikeda: "Aspergillus species strain M39 produces two naphthoーγーpyrones that reverse drug resistance in human KB cells." International Journal of Cancer. 45. 508-513 (1990)
[Publications] Yasuo Kuwazuru: "Expression of the multidrug transporter,Pーglycoprotein,in acute leukemia cells and correlation to clinical drug resistance." Cancer. 66. 868-873 (1990)
[Publications] Misako Ichikawa: "Modulators of the multidrug transporter,Pーglycoprotein,exist in the human plasma." Biochemical Biophysical Research Communication. 166. 74-80 (1990)
[Publications] Norimasa Shudo: "Two pyridine analogues with more effective ability to reverse multidrug resistance and with lower calcium channel blocking activity than their dihydropyridine counterparts." Cancer Research. 50. 3055-3061 (1990)
[Publications] Yasuo Kuwazuru: "Expression of Pーglycoprotein in adult T cell leukemia cells." Blood. 76. 2065-2071 (1990)
[Publications] Tetsuro Mizoguchi: "Expression of the MDR1 gene in human gastric and colorectal carcinoma." Journal of National Cancer Institute. 82. 1679-1683 (1990)
[Publications] Misako Ichikawa: "Interaction of organic chemicals with Pーglycoprotein in the adrenal gland,kidney,and multidrugーresistant KB cell." Journal of Biological Chemistry. 266. 903-908 (1991)
[Publications] 秋山伸一: "抗癌剤のはたらきー課題と展望ー" 共立出版株式会社, 1-130 (1990)
[Publications] Shinーichi Akiyama: "HeLa Cells in “Encyclopedia of Human Biology,Vol.4"" Academic Press, 1-5 (1990)
[Publications] Shinーichi Akiyama: "A molecular basis for multidrugーresistance and reversal of the resistance in Proceedings of the 8th Nikko Brain Tumor Conference" SpringerーVerlag,Tokyo,Inc., (1991)

1990 Fiscal Year Annual Research Report

Pー糖蛋白質の構造と機能の研究

Principal Investigator

秋山 伸一 鹿児島大学, 医学部, 教授 (60117413)

Research Products

[Publications] Yasuo Kuwazuru: "Expression of the multidrug transporter,Pーglycoprotein,in chronic myelogenous leukemia cells in blast crisis." British Journal of Haematology. 74. 24-29 (1990)

[Publications] Akihiko Yoshimura: "Novel screening method for agents that overcome classical multidrug resistance in a human cell line." Cancer Letters. 50. 45-51 (1990)

[Publications] Akihiko Yoshimura: "Purification and tissue distribution of human thymidine phosphorylase;high expression in lymphocytes,reticulocytes and tumors." Biochimica Biophysica Acta. 1034. 107-113 (1990)

[Publications] Shunーichi Ikeda: "Aspergillus species strain M39 produces two naphthoーγーpyrones that reverse drug resistance in human KB cells." International Journal of Cancer. 45. 508-513 (1990)

[Publications] Yasuo Kuwazuru: "Expression of the multidrug transporter,Pーglycoprotein,in acute leukemia cells and correlation to clinical drug resistance." Cancer. 66. 868-873 (1990)

[Publications] Misako Ichikawa: "Modulators of the multidrug transporter,Pーglycoprotein,exist in the human plasma." Biochemical Biophysical Research Communication. 166. 74-80 (1990)

[Publications] Norimasa Shudo: "Two pyridine analogues with more effective ability to reverse multidrug resistance and with lower calcium channel blocking activity than their dihydropyridine counterparts." Cancer Research. 50. 3055-3061 (1990)

[Publications] Yasuo Kuwazuru: "Expression of Pーglycoprotein in adult T cell leukemia cells." Blood. 76. 2065-2071 (1990)

[Publications] Tetsuro Mizoguchi: "Expression of the MDR1 gene in human gastric and colorectal carcinoma." Journal of National Cancer Institute. 82. 1679-1683 (1990)

[Publications] Misako Ichikawa: "Interaction of organic chemicals with Pーglycoprotein in the adrenal gland,kidney,and multidrugーresistant KB cell." Journal of Biological Chemistry. 266. 903-908 (1991)

[Publications] 秋山 伸一: "抗癌剤のはたらきー課題と展望ー" 共立出版株式会社, 1-130 (1990)

[Publications] Shinーichi Akiyama: "HeLa Cells in “Encyclopedia of Human Biology,Vol.4"" Academic Press, 1-5 (1990)

[Publications] Shinーichi Akiyama: "A molecular basis for multidrugーresistance and reversal of the resistance in Proceedings of the 8th Nikko Brain Tumor Conference" SpringerーVerlag,Tokyo,Inc., (1991)

秋山伸一鹿児島大学, 医学部, 教授 (60117413)

[Publications] 秋山伸一: "抗癌剤のはたらきー課題と展望ー" 共立出版株式会社, 1-130 (1990)