1990 年度実績報告書

チトクロ-ムPー450の分子生物学・機能の分子機構

研究課題

研究課題/領域番号	02217103
研究機関	慶応義塾大学
研究代表者	石村巽慶應義塾大学, 医学部, 教授 (40025599)
研究分担者	岡本光弘大阪大学, 医学部, 教授 (90028613) 武森重樹広島大学, 総合科学部, 教授 (60019461) 北川禎三岡崎国立共同研究機構, 分子科学研究所, 教授 (40029955) 川戸佳東京大学, 教養学部, 助教授 (50169736) 森島績京都大学, 工学部, 教授 (50026093)
キーワード	チトクロ-ムPー450 / Pー450_<11β> / Pー450_<aldo> / Pー450_<14DM> / 副腎皮質ミトコンドリア / Pー450_<cam> / 部位特異的変異
研究概要	1.石村らはヒトのアルドステロン合成酵素(Pー450aldo)をアルドステロン産生腫瘍から精製・分離し(63回日本生化学会発表、J.Biol.Chem.in press)、これがCYP11B2遺伝子の産物であることを示した。またラット副腎皮質球状層におけるPー450aldoの発現がアンギオテンシンIIおよびK^+により調節されることを証明した(63回日本内分泌学会発表、Endocrinology,in press)。さらに岡本らはラットPー450aldoのcDNAをCOS細胞に発現させ、そのアルドステロン合成活性を確認した(Biochem.Biophys.Res.Commun.169他)。以上によりアルドステロン合成酵素の実体と分子的性質が解明された。一方、武森らはウシ副腎皮質培養細胞系等を用いてPー450 _<17α,lyase>ならびにPー450_<c21>が関与するミクロソ-ム電子伝達系の反応を解析し、ステロイド代謝の調節機構を明らかにした(J.Steroid.Biochem.投稿中)。井柳らはPー450の酸化反応に共役するグルクロン酸転移酵素の遺伝子解析を行なった(63回日本生化学会発表)。 2.北川、飯塚、森島らはそれぞれ共鳴ラマン散乱、X線吸収スペクトル、二次元NMR法を用いてP450camの構造ー活性相関を解明し、川戸らはフラッシュホトリシス偏光解消法を用いて膜中の各種Pー450の回転運動と蛋白質相互作用を測定した(J.Biochem.107他)。吉田らは酵母Pー450_<14DM>(ラノステロ-ル14αー脱メチル化酵素)の活性中心と其質の相互作用を各種のアナログを用いて解析した(Chirality,2他)。

研究成果

(25件)

すべてその他

すべて文献書誌 (25件)

[文献書誌] Imai,M.: "Molecular Cloning of a cDNA Encoding Aldosterone Synthase Cytochrome Pー450 in Rat Adrenal Cortex" FEBS Letters. 263. 299-302 (1990)
[文献書誌] Shibata,H.: "Regulation of Aldosterone Synthase Cytochrome Pー450 in Rat Adrenals by Angiotensin II and Potassuium" Endocrinology. 128. (1991)
[文献書誌] Ogishima,T.: "Aldosterone Synthase Cytochrome Pー450 expressed in the Adrenals of Patients with Primary Aldosteronism" The Journal of Biological Chemistry.(1991)
[文献書誌] Sato,H.: "Thermodynamic Study of Protein Dynamic Structure in the Oxygen Binding Reaction of Myoglobin" The Journal of Biological Chemistry. 265. 18823-18828 (1990)
[文献書誌] Kamiya,N.: "Heme Environmental Structure of Novel Artificial Myoglobin with Closed Heme Pocket:SiteーSpecific ChemicalーModification Producing Distal NーTetrazolyーHistydine E7 by Cyanogen Bromide and Azide Ion." The Journal of the American Chemistry. (1991)
[文献書誌] Ueno,I.: "Characterization of bーType Cytochrome in situ by Electron Paramagnetic Resonance Spectroscopy" FEBS Letters. (1991)
[文献書誌] Unno,M.: "High Pressure Laser Photolysis Study of Hemoproteins.Effects of Pressure on Carbon Monoxide Binding Dymamics for Rーand TーState Hemoglobins." Biochemistry. 29. 10199-10205 (1990)
[文献書誌] Nakashima,S.: "Spin Coupling in Ferric Porphyrin and Chlorin πーCation Radical Complexes." Inorganic Chemistry. 29. 5207-5211 (1990)
[文献書誌] Unno,M.: "Pressure Effects on the Carbon Monoxide Rebinding to the Isolated α and β Chains of Human Hemoglobin." The Jounal of the American Chemical Society.
[文献書誌] Egawa,T.: "Resonance Raman Study on Mutant Cytochrome Pー450 Obtained by Site Directed Mutagenesis." Biochimica et Biophysica Acta. 1040. 211-216 (1990)
[文献書誌] Ogura,T.: "Observation of the Fe^<4+>=O Stretching Raman Band for Cytochrome Oxidase Compound B at Ambient Temperature" The Journal of Biological Chemistry. 265. 14721-14723 (1990)
[文献書誌] Egawa,T.: "Observation the OーO Stretching Raman Band for Cytochrome Pー450_<cam> under Catalytic Conditions." The Journal of Biological Chemistry. (1991)
[文献書誌] Ohta,Y.: "Conversion of Cholesterol to Pregnenolone Mobilizes Cytochrome Pー450 in the Inner Membrane of Adrenocortical Mitochondria:Protein Rotation Study" The Journal of Biochemistry. 107. 97-104 (1990)
[文献書誌] Soma,Y.: "Effects of Oxygen on the Relative Photodissociability of Cytochrome Pー450・CO Complex in Rat Liver Microsomes." Biochemistry. (1991)
[文献書誌] Kawato,S.: "DrugーInduction Decreases the Mobility of Cytochrome Pー450 in Rat Liver Microsomes:Protein Rotation Study" The Journal of Biochemistry. (1991)
[文献書誌] Iyanagi,T.: "Molecular Basis of Multiple UDPーGlucuronosyltransferase Isoenzyme Deficiencies in the Hyperbilirubinemic Rat(Gunn Rat)" The Journal of Biological Chemistry. (1991)
[文献書誌] Shimura,K.: "Immunocytochemical and Biochemical Studies on the Localization and Changes of 17αーHydroxylase/C17ーC20 lyase Activity in Immature Rat Ovary Treated with PMSG and hCG" Histochemistry. 94. 225-229 (1990)
[文献書誌] Yamazaki,T.: "Cytochrome Pー450(17α,lyase)ーMediating Pathway of Androgen Synthesis in Bovine Adrenocortical Cultured Cells." The Journal of Steroid Biochemistry. (1990)
[文献書誌] Kominami,S.: "The Role of Cytochrome b5 in Adrenal Microsomal Steroidogenesis" The Journal of Steroid Biochemistry. (1991)
[文献書誌] Morohashi,K.: "Enzymatic Activities of Pー450(11β)s Expressed by Two cDNAs in COSー7 Cells." The Journal of Biochemistry. 107. 635-640 (1990)
[文献書誌] Matsukawa,N.: "Molecular Cloning and Expression of cDNAs Encoding Rat Aldosterone Synthase,Variants of Cytochrome Pー450_<11β>" Biochemical and Biophysical Research Communications. 169. 245-252 (1990)
[文献書誌] Nonaka,Y.: "Molecular Nature of Aldosterone Synthase,a Member of Cytochrome Pー450_<11β> Family" Endocrine Research. (1991)
[文献書誌] Kaergel,E.: "Comparative Study on Cytochrome Pー450 of Yeast Using Specific Antibodies to Cytochrome Pー450_<alk> and Pー450_<14DM>" Yeast. 6. 61-67 (1990)
[文献書誌] Yoshida,Y.: "Stereoselective Interaction of an Azole Antifungal Agent with Its Target,Lanosterol 14αーDemethylase(Cytochrome Pー450_<14DM>):A Model Study with Stereoisomers of Triadimenol and Purified Cytochrome Pー450_<14DM> from Yeast" Chirality. 2. 10-15 (1990)
[文献書誌] Aoyama,Y.: "Role of the Side Chain of Lanosterol in Substrate Recognition and Chatalytic Activity of Lanosterol 14αーDemethylase(Cytochrome Pー450_<14DM>)of Yeast" Biochimica et Biophysica Acta. (1991)

1990 年度 実績報告書

チトクロ-ムPー450の分子生物学・機能の分子機構

研究代表者

石村 巽 慶應義塾大学, 医学部, 教授 (40025599)

研究成果

[文献書誌] Imai,M.: "Molecular Cloning of a cDNA Encoding Aldosterone Synthase Cytochrome Pー450 in Rat Adrenal Cortex" FEBS Letters. 263. 299-302 (1990)

[文献書誌] Shibata,H.: "Regulation of Aldosterone Synthase Cytochrome Pー450 in Rat Adrenals by Angiotensin II and Potassuium" Endocrinology. 128. (1991)

[文献書誌] Ogishima,T.: "Aldosterone Synthase Cytochrome Pー450 expressed in the Adrenals of Patients with Primary Aldosteronism" The Journal of Biological Chemistry.(1991)

[文献書誌] Sato,H.: "Thermodynamic Study of Protein Dynamic Structure in the Oxygen Binding Reaction of Myoglobin" The Journal of Biological Chemistry. 265. 18823-18828 (1990)

[文献書誌] Kamiya,N.: "Heme Environmental Structure of Novel Artificial Myoglobin with Closed Heme Pocket:SiteーSpecific ChemicalーModification Producing Distal NーTetrazolyーHistydine E7 by Cyanogen Bromide and Azide Ion." The Journal of the American Chemistry. (1991)

[文献書誌] Ueno,I.: "Characterization of bーType Cytochrome in situ by Electron Paramagnetic Resonance Spectroscopy" FEBS Letters. (1991)

[文献書誌] Unno,M.: "High Pressure Laser Photolysis Study of Hemoproteins.Effects of Pressure on Carbon Monoxide Binding Dymamics for Rーand TーState Hemoglobins." Biochemistry. 29. 10199-10205 (1990)

[文献書誌] Nakashima,S.: "Spin Coupling in Ferric Porphyrin and Chlorin πーCation Radical Complexes." Inorganic Chemistry. 29. 5207-5211 (1990)

[文献書誌] Unno,M.: "Pressure Effects on the Carbon Monoxide Rebinding to the Isolated α and β Chains of Human Hemoglobin." The Jounal of the American Chemical Society.

[文献書誌] Egawa,T.: "Resonance Raman Study on Mutant Cytochrome Pー450 Obtained by Site Directed Mutagenesis." Biochimica et Biophysica Acta. 1040. 211-216 (1990)

[文献書誌] Ogura,T.: "Observation of the Fe^<4+>=O Stretching Raman Band for Cytochrome Oxidase Compound B at Ambient Temperature" The Journal of Biological Chemistry. 265. 14721-14723 (1990)

[文献書誌] Egawa,T.: "Observation the OーO Stretching Raman Band for Cytochrome Pー450_<cam> under Catalytic Conditions." The Journal of Biological Chemistry. (1991)

[文献書誌] Ohta,Y.: "Conversion of Cholesterol to Pregnenolone Mobilizes Cytochrome Pー450 in the Inner Membrane of Adrenocortical Mitochondria:Protein Rotation Study" The Journal of Biochemistry. 107. 97-104 (1990)

[文献書誌] Soma,Y.: "Effects of Oxygen on the Relative Photodissociability of Cytochrome Pー450・CO Complex in Rat Liver Microsomes." Biochemistry. (1991)

[文献書誌] Kawato,S.: "DrugーInduction Decreases the Mobility of Cytochrome Pー450 in Rat Liver Microsomes:Protein Rotation Study" The Journal of Biochemistry. (1991)

[文献書誌] Iyanagi,T.: "Molecular Basis of Multiple UDPーGlucuronosyltransferase Isoenzyme Deficiencies in the Hyperbilirubinemic Rat(Gunn Rat)" The Journal of Biological Chemistry. (1991)

[文献書誌] Shimura,K.: "Immunocytochemical and Biochemical Studies on the Localization and Changes of 17αーHydroxylase/C17ーC20 lyase Activity in Immature Rat Ovary Treated with PMSG and hCG" Histochemistry. 94. 225-229 (1990)

[文献書誌] Yamazaki,T.: "Cytochrome Pー450(17α,lyase)ーMediating Pathway of Androgen Synthesis in Bovine Adrenocortical Cultured Cells." The Journal of Steroid Biochemistry. (1990)

[文献書誌] Kominami,S.: "The Role of Cytochrome b5 in Adrenal Microsomal Steroidogenesis" The Journal of Steroid Biochemistry. (1991)

[文献書誌] Morohashi,K.: "Enzymatic Activities of Pー450(11β)s Expressed by Two cDNAs in COSー7 Cells." The Journal of Biochemistry. 107. 635-640 (1990)

[文献書誌] Matsukawa,N.: "Molecular Cloning and Expression of cDNAs Encoding Rat Aldosterone Synthase,Variants of Cytochrome Pー450_<11β>" Biochemical and Biophysical Research Communications. 169. 245-252 (1990)

[文献書誌] Nonaka,Y.: "Molecular Nature of Aldosterone Synthase,a Member of Cytochrome Pー450_<11β> Family" Endocrine Research. (1991)

[文献書誌] Kaergel,E.: "Comparative Study on Cytochrome Pー450 of Yeast Using Specific Antibodies to Cytochrome Pー450_<alk> and Pー450_<14DM>" Yeast. 6. 61-67 (1990)

[文献書誌] Yoshida,Y.: "Stereoselective Interaction of an Azole Antifungal Agent with Its Target,Lanosterol 14αーDemethylase(Cytochrome Pー450_<14DM>):A Model Study with Stereoisomers of Triadimenol and Purified Cytochrome Pー450_<14DM> from Yeast" Chirality. 2. 10-15 (1990)

[文献書誌] Aoyama,Y.: "Role of the Side Chain of Lanosterol in Substrate Recognition and Chatalytic Activity of Lanosterol 14αーDemethylase(Cytochrome Pー450_<14DM>)of Yeast" Biochimica et Biophysica Acta. (1991)

1990 年度実績報告書

石村巽慶應義塾大学, 医学部, 教授 (40025599)