1989 年度実績報告書

チトクローム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	01635004
研究機関	慶応義塾大学
研究代表者	石村巽慶応義塾大学, 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) 片桐正之金沢大学, 理学部, 教授 (30019442) 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	チトクロームPー450 / P45011β / Pー450aldo / Pー450_<14DM> / 副腎ミトコンドリア / Pー450cam / 部位特異的変異
研究概要	1)石村らはPー450cam遺伝子に部位特異的変異を導入して、カンファーの水酸化反応にはPー450camの酸素結合部位近傍にあるスレオニンー252がプロトン供与体として重要な役割を果たしていることを明らかにした(Pro.Nat.Acad.Sci.86)。一方、昨年その単離精製に成功したPー450の新しい分子種であるアルドステロン合成酵素(Pー450aldo)はNa欠食ラットの副腎球状層に著しく増加することを見出した(J,Biol.Chem,264)。2)森島ら、飯塚らは高分解能NMR、閃光分解法、更にXAFS法などを用いてP450における配位子結合とそれに伴う微細な構造変化との関係を解明した(Biochem.28 他)。3)北川らは遺伝子操作により調整したPー450ωー1の変異酵素を用いてスレオニンー301がヘム近傍にあることをFeーCO伸縮振動の測定から推測した(Biochem.28 他)。4)川戸らはPー450・CO複合体のフラッシュフオトリシス偏光解消を利用してミトコンドリア及びミクロゾーム膜中におけるPー450の回転運動を測定し、Pー450はその電子伝達成分と一時的な複合体を形成することを明らかにした(J,Biochem.107 他)。5)片桐ら、武森らはPー450llβの多様な触媒作用を明きらかにし、特にウシ副腎皮質各層ではミトコンドリア内膜レベルでPー450llβへの電子伝達が調節を受けており、球状層では電子が多重に流れて連続的な水酸化反応がおこりアルドステロンが特異的に生成されている可能性を示唆した(Biochem.Biophys.Acta.984 他)。6)岡本らは、Pー450llβのアルドステロン合成能を解明する目的でラットPー450llβのcDNAをクローン化し、その塩基配列の決定とCOS細胞内での活性発現を行った。(FEBS.Lett.255 他)7)吉田らは基質構造類似体を用いてPー450_<14DM>と基質との相互作用を系統的に解析し、反応に必須な基質の基本構造を推定した(Biochem.Biophys.Acta.1006 他)。

研究成果
(27件)

すべてその他

すべて文献書誌 (27件)

[文献書誌] Imai M.: "Uncoupling of the Cytochrome Pー450cam Monooxygenese Reaction by a Single Mutation of Threonine 252 to Alanine or Valine:A Possible Role of the Hydroxy Amino Acid in the Oxygen Activation" Proceedings of National Academy of Sciences. 86. 7823-7827 (1989)
[文献書誌] Nakamura Y.: "Superozxide Anion is the Initial Product in the Hydrogen Peroxide Formation Catalyzed by NADPHーOxidase in Porcine Thyroid Plasma" The Journal of Biological Chemistry. 264. 4759-4761 (1989)
[文献書誌] Ogishima T.: "Isolation of Aldosterone Synthase Cytochrome Pー450 from Zona Glomerulosa Mitochondria of Rat Adrenal Cortex" The Journal of Biological Chemistry. 264. 10935-10938 (1989)
[文献書誌] Watanabe T.: "A Model Study on Aromatase Cytochrome Pー450 Reaction:Transformation of Androstenー19ーalー3,17ーdione to 10βーHydroxyest γー4ーene3,17ーdione" The Journal of the American Chemical Society. 111. 8047-8049 (1989)
[文献書誌] Shiro Y.: "^<15>NーNMR Study on Cyanide(C^<15>N^-)Complex of Cytochrome Pー450cam.Effects of DーCamphor and Putidaredoxin on IronーLigand Structure" The Journal of the American Chemical Society. 111. 7707-7711 (1989)
[文献書誌] Sato H.: "New Transient Species of Sperm Whale Myogiobin in Photodissociation of Dioxygen from Oxymyoglobin" The Jounal of Biological Chemistory. (1990)
[文献書誌] Shiro Y.: "XーRay Absorption Spectral Studies of Ferric High Spin Hemoproteins:XANES Evidences for Coordination Structure of the Heme iron" The Journal of American Chemical Society. (1990)
[文献書誌] Ishimori K.: "NMR Study of Human Mutant Hemoglobins Synthesized in E Coli." The Journal of Biological Chemistry. 264. 14624-14626 (1989)
[文献書誌] Adachi S.: "The Effects of Pressure on Oxygen and Carbon Monoxide Binding Kinetics for Myoglobin" The Journal of Biological Chemistry. 264. 18896-18901 (1989)
[文献書誌] Morishima I.: "Effect of the Distal Histidine Modification(cyanation)of Myoglobin on the Ligand Binding Kinetics and the Heme Environmental Structures" Biochemistry. 28. 7582-7586 (1989)
[文献書誌] Nakagawa M.: "Origin of Raman Spectral Difference between the Spinningーcell and Flowーcell Measurements for Bacteriorhodopsin Photointermediates" Journal of Raman Spectroscopy. 20. 303-305 (1989)
[文献書誌] Nagai M.: "Caracteristics in Tyroshine Coordination of four Hemoglobins M probed by Resonance Raman Spectroscopy" Biochemistry. 28. 2418-2422 (1989)
[文献書誌] Ogura T.: "Raman/Absorption Simultaneous Measurements for Cytochrome Oxidase Compound A at Room Temperature with a Novel Flow Aparatus" Biochemistry. 28. 8022-8027 (1989)
[文献書誌] Iyanagi T.: "The Methylcholanthreneーinducible UDPーGlucuronyltransferase Deficiency in the Hyperbilirubinemic Rat(Gunn Rat)is caused by a-1 Frameshift Mutation" The Journal of Biological Chemistry. 264. 21302-21307 (1989)
[文献書誌] Iyanagi T.: "On the Mechanism of OneーElectron Reduction of Quinoes by Microsomal Flavin Enzymes:The Kinetic Analysis between Cytochrome b_5 and Menadione" Free Radical Research Communications. (1989)
[文献書誌] Suzuki S.: "Independent Flexible Motion of Submolecular Domains of the Ca^<2+>,Mg^<2+>ーATPase of Sarcoplasmic Reticulum Measured by TimeーResolved Fluorescence Depolarization of SiteーSpecifically Attached Probes" Biochemistry. 28. 7734-7740 (1989)
[文献書誌] Ohta Y.: "Conversi on of Cholesterol to Pregnenolone Mobilizes Cytochrome Pー450in the Inner Membrane of Adrenocortical Mitochondria:Protein Rotation Study" 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. (1990)
[文献書誌] Ikusiro S.: "Adrenal Cytochrome Pー45011βーproteoliposomes Catalyzing Aldosteron Synthesis:Preparation and Characterization" Biochimica et Biophysica Acta. 984. 50-56 (1989)
[文献書誌] Kominami S.: "Steroidogenesis in Liposomal System Containing Adrenal Microsomal Cytochrome Pー450 Electron Transfer Components" Biochimicaet Biophysica Acta. 985. 293-299 (1989)
[文献書誌] Kominami S.: "Steroidogenesis in Liposomal System Containing Adrenal Microsomal Electron Transfer Components" Cytochrome Pー450:Biochemistry and Biophysics(ed.I.Schuster)Taylor and Francis,London. 223-226 (1989)
[文献書誌] Okamoto M.: "Cytochrome Pー45011βーmediated 19ーOxidation Pathway of 11ーDeoxy corticosterone" The Adrenal and Hypertension:From Cloning to Clinic.Sereno Symposia Publications(eds.F.Mantero et al)Raven Press,New York. 57. 439-444 (1989)
[文献書誌] Sasano H.: "Immunolocalization of Aromatase,17αーHydroxylase and SideーChain Cleavage Cytochromeー450 in Human Ovary" Journal of Reproduction and Fertility. 85. 163-169 (1989)
[文献書誌] Nonaka Y.: "Molecular Cloning and Sequence Analysis of cDNA Encoding Rat Adrenal Cytochrome Pー45011β" FEBS Letters. 255. 21-26 (1989)
[文献書誌] Aoyama Y.: "The 3ーHydroxy Group of Lanosterol is Essential for Originating the Substrate in the Substrate Site of Cytochrome Pー450_<14DM>(Lanosterol 14αーdemethylase)" Biochemica et Biophysica Acta. 1006. 209-213 (1989)
[文献書誌] Aoyama Y.: "Deformylation of 32ーO_<xo>ー24,25ーdihydrolanosterrol by the Purified Cytochrome Pー450_<14DM>(Lanosterol 14αーdemethylase)from yeast.Evidence Confirming the Intermediate Step of Lanosterol 14αーDeーmethylation" The Journal of Biological Chemistry. 264. 18502-18505 (1989)
[文献書誌] Yoshida Y.: "Stereoselective Interaction of an Azole Antifungal Agent with its Target,Lanosterol 14αーDemethylase(CytochromePー450_<14DM>):A model study with Stereoisomers Triadimenol and Purified Cytochrome P-450_<14DM> From Yeast." Chirality. (1990)

1989 年度 実績報告書

チトクロームPー450の分子生物学:機能の分子機構

研究代表者

石村 巽 慶応義塾大学, 医学部, 教授 (40025599)

研究成果

[文献書誌] Imai M.: "Uncoupling of the Cytochrome Pー450cam Monooxygenese Reaction by a Single Mutation of Threonine 252 to Alanine or Valine:A Possible Role of the Hydroxy Amino Acid in the Oxygen Activation" Proceedings of National Academy of Sciences. 86. 7823-7827 (1989)

[文献書誌] Nakamura Y.: "Superozxide Anion is the Initial Product in the Hydrogen Peroxide Formation Catalyzed by NADPHーOxidase in Porcine Thyroid Plasma" The Journal of Biological Chemistry. 264. 4759-4761 (1989)

[文献書誌] Ogishima T.: "Isolation of Aldosterone Synthase Cytochrome Pー450 from Zona Glomerulosa Mitochondria of Rat Adrenal Cortex" The Journal of Biological Chemistry. 264. 10935-10938 (1989)

[文献書誌] Watanabe T.: "A Model Study on Aromatase Cytochrome Pー450 Reaction:Transformation of Androstenー19ーalー3,17ーdione to 10βーHydroxyest γー4ーene3,17ーdione" The Journal of the American Chemical Society. 111. 8047-8049 (1989)

[文献書誌] Shiro Y.: "^<15>NーNMR Study on Cyanide(C^<15>N^-)Complex of Cytochrome Pー450cam.Effects of DーCamphor and Putidaredoxin on IronーLigand Structure" The Journal of the American Chemical Society. 111. 7707-7711 (1989)

[文献書誌] Sato H.: "New Transient Species of Sperm Whale Myogiobin in Photodissociation of Dioxygen from Oxymyoglobin" The Jounal of Biological Chemistory. (1990)

[文献書誌] Shiro Y.: "XーRay Absorption Spectral Studies of Ferric High Spin Hemoproteins:XANES Evidences for Coordination Structure of the Heme iron" The Journal of American Chemical Society. (1990)

[文献書誌] Ishimori K.: "NMR Study of Human Mutant Hemoglobins Synthesized in E Coli." The Journal of Biological Chemistry. 264. 14624-14626 (1989)

[文献書誌] Adachi S.: "The Effects of Pressure on Oxygen and Carbon Monoxide Binding Kinetics for Myoglobin" The Journal of Biological Chemistry. 264. 18896-18901 (1989)

[文献書誌] Morishima I.: "Effect of the Distal Histidine Modification(cyanation)of Myoglobin on the Ligand Binding Kinetics and the Heme Environmental Structures" Biochemistry. 28. 7582-7586 (1989)

[文献書誌] Nakagawa M.: "Origin of Raman Spectral Difference between the Spinningーcell and Flowーcell Measurements for Bacteriorhodopsin Photointermediates" Journal of Raman Spectroscopy. 20. 303-305 (1989)

[文献書誌] Nagai M.: "Caracteristics in Tyroshine Coordination of four Hemoglobins M probed by Resonance Raman Spectroscopy" Biochemistry. 28. 2418-2422 (1989)

[文献書誌] Ogura T.: "Raman/Absorption Simultaneous Measurements for Cytochrome Oxidase Compound A at Room Temperature with a Novel Flow Aparatus" Biochemistry. 28. 8022-8027 (1989)

[文献書誌] Iyanagi T.: "The Methylcholanthreneーinducible UDPーGlucuronyltransferase Deficiency in the Hyperbilirubinemic Rat(Gunn Rat)is caused by a-1 Frameshift Mutation" The Journal of Biological Chemistry. 264. 21302-21307 (1989)

[文献書誌] Iyanagi T.: "On the Mechanism of OneーElectron Reduction of Quinoes by Microsomal Flavin Enzymes:The Kinetic Analysis between Cytochrome b_5 and Menadione" Free Radical Research Communications. (1989)

[文献書誌] Suzuki S.: "Independent Flexible Motion of Submolecular Domains of the Ca^<2+>,Mg^<2+>ーATPase of Sarcoplasmic Reticulum Measured by TimeーResolved Fluorescence Depolarization of SiteーSpecifically Attached Probes" Biochemistry. 28. 7734-7740 (1989)

[文献書誌] Ohta Y.: "Conversi on of Cholesterol to Pregnenolone Mobilizes Cytochrome Pー450in the Inner Membrane of Adrenocortical Mitochondria:Protein Rotation Study" 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. (1990)

[文献書誌] Ikusiro S.: "Adrenal Cytochrome Pー45011βーproteoliposomes Catalyzing Aldosteron Synthesis:Preparation and Characterization" Biochimica et Biophysica Acta. 984. 50-56 (1989)

[文献書誌] Kominami S.: "Steroidogenesis in Liposomal System Containing Adrenal Microsomal Cytochrome Pー450 Electron Transfer Components" Biochimicaet Biophysica Acta. 985. 293-299 (1989)

[文献書誌] Kominami S.: "Steroidogenesis in Liposomal System Containing Adrenal Microsomal Electron Transfer Components" Cytochrome Pー450:Biochemistry and Biophysics(ed.I.Schuster)Taylor and Francis,London. 223-226 (1989)

[文献書誌] Okamoto M.: "Cytochrome Pー45011βーmediated 19ーOxidation Pathway of 11ーDeoxy corticosterone" The Adrenal and Hypertension:From Cloning to Clinic.Sereno Symposia Publications(eds.F.Mantero et al)Raven Press,New York. 57. 439-444 (1989)

[文献書誌] Sasano H.: "Immunolocalization of Aromatase,17αーHydroxylase and SideーChain Cleavage Cytochromeー450 in Human Ovary" Journal of Reproduction and Fertility. 85. 163-169 (1989)

[文献書誌] Nonaka Y.: "Molecular Cloning and Sequence Analysis of cDNA Encoding Rat Adrenal Cytochrome Pー45011β" FEBS Letters. 255. 21-26 (1989)

[文献書誌] Aoyama Y.: "The 3ーHydroxy Group of Lanosterol is Essential for Originating the Substrate in the Substrate Site of Cytochrome Pー450_<14DM>(Lanosterol 14αーdemethylase)" Biochemica et Biophysica Acta. 1006. 209-213 (1989)

[文献書誌] Yoshida Y.: "Stereoselective Interaction of an Azole Antifungal Agent with its Target,Lanosterol 14αーDemethylase(CytochromePー450_<14DM>):A model study with Stereoisomers Triadimenol and Purified Cytochrome P-450_<14DM> From Yeast." Chirality. (1990)

1989 年度実績報告書

石村巽慶応義塾大学, 医学部, 教授 (40025599)