1990 Fiscal Year Annual Research Report

チトクロ-ムPー450の分子生物学:発現の分子機構

Research Project

Project/Area Number	02217102
Research Institution	Kyushu University
Principal Investigator	大村恒雄九州大学, 大学院医学系研究科, 教授 (80029933)
Co-Investigator(Kenkyū-buntansha)	渡辺民朗東北大学, 抗酸菌病研究所, 教授 (40006101) 堀内忠郎九州大学, 薬学部, 教授 (10037567) 藤井義明東北大学, 理学部, 教授 (00098146) 加藤隆一慶応義塾大学, 医学部, 教授 (40112685) 奥田九一郎広島大学, 歯学部, 教授 (80034156)
Keywords	チトクロ-ムPー450 / Pー450 / 発現の分子機構 / 発現調節
Research Abstract	新たなPー450分子種の確認と構造決定、Pー450遺伝子の発現調節機構、Pー450の生合成と膜への組み込み機構、Pー450の構造と酵素活性の相関などの諸問題を研究班員が分担して研究を進めた。新たなPー450として、ラット肝のコレステロ-ル7α位を水酸化するPー450(奥田)、ラット腎臓の25ーヒドロキシビタミンD_3の24位を水酸化するPー450(奥田)、ラット肝細胞の初代培養で誘導されるPー450(大村)など数種類の一次構造を決定した。 Pー450遺伝子の発現調節については、ラット肝臓においてのPー450aの発現が成長ホルモンにより調節されることを確かめ(加藤)、Pー450c遺伝子のTCDD誘導時に発現制御領域XREに結合する因子の精製を進め(船江)、BTEに働く新しい因子の構造を決定した(藤井)。また、ステロイドホルモン生合成に関与するPー450_<11>β(大村)、Pー450sccとPー450c_<21>(藤井)の転写調節領域を解析し、新しいエレメントの存在を確認した。ヒトについては、Pー450c遺伝子の多型と肺癌発生の間に有意の相関があることを見いだし(川尻)、胎児型Pー450の発現も検討した(鎌滝)。又、ハムスタ-の肺Pー450についても発現調節の研究を進めた(渡辺)。 Pー450の生合成については、Pー450が生合成時に小胞体膜へ組み込まれる過程を解析し、組み込み時にシグナルとして機能するアミノ酸配列の構造要求性を検討した(大村)。また、肝細胞内においてのPー450の分解過程を免疫電顕法で解析した(田代)。 Pー450の構造活性相関については、Pー450cam(堀内)、Pー450d(清水)、及びPー450IIC(今井)についてアミノ酸点変異の導入、キメラ蛋白質の作成などにより研究を進め、活性に必要ないくつかのアミノ酸残基を特定できた。

Research Products
(16 results)

All Other

All Publications (16 results)

[Publications] T.Sato: "The Aminoーterminal Structures That Determine Topological Orientation of Cytochrome Pー450 in Microsomal Membrane" EMBO J.9. 2391-2397 (1990)
[Publications] Shinーichirou Honda: "Novel cAMP Regulatory Elements in the Promotor Region of Bovine Pー450(11β) Gene" J.Biochem.108. 1042-1049 (1990)
[Publications] Egawa,T: "Resonance Raman Study on Mutant Cytochrome Pー450 Obtained by SiteーDirected Mutagenesis" Biochim.Biophys.Acta. 1040. 211-216 (1990)
[Publications] Imai,Y: "Characterization of Rabbit Liver P450IIE1 Synthesized in Transformed Yeast Cells" J.Biochem.108. 522-524 (1990)
[Publications] M.Noshiro: "Rat Liver Cholesterol 7αーHydroxylase" J.Biol.Chem.265. 10036-10041 (1990)
[Publications] Yamazoe,Y: "Modulation of hepatic level of microsomal testosterone 7αーhydroxylase,Pー450a(Pー450IIA),by thyroid hormone and growth hormone in rat liver" J.Biochem.108. 599-603 (1990)
[Publications] Nagata,K: "Purification and characterization of four catalytically active testosterone 6βーhydroxylase Pー450s from rat liver microsomes: comparison of a novel form with three structurally and functionally related forms" J.Biochem.107. 718-725 (1990)
[Publications] M.Kitada: "Metabolic activation of aflatoxin B,and 2ーaminoー3ーmethylimidazo(4,5ーf)quinoline by human adult and fetal livers" Cancer Res.50. 2641-2645 (1990)
[Publications] T.Uchida: "Isolation of cDNA coding for three different forms of liver microsomal cytochrome Pー450 from polychlorinated biphenylーtreated beagle dogs" Mol.Pharmacol.38. 644-651 (1990)
[Publications] H.Sotokawa: "Electron Spin Resonance Studies of WildーType and Mutant Cytochromes Pー450_d: Effects of Mutations at Proximal,Aromatic and Distal Sites on g__ー Values" Biochim.Biophys.Acta. 1037. 122-128 (1990)
[Publications] Yamamoto,A: "Absence of cytochrome Pー450 and presence of autolysosomal membrane antigens on the isolation membranes and autophagosomal membranes in rat hepatocytes" J.Histochem.Cytochem.38. 1571-1581 (1990)
[Publications] Yokotani,N: "cDNA cloning of cytochrome Pー450 related to Pー450_pー2 from the expression" Eur.J.Biochem.187. 23-29 (1990)
[Publications] Yanagida,A: "A novel cisーacting DNA element required for a high level of inducible expression of the rat Pー450c gene" Mol.Cell.Biol.10. 1470-1475 (1990)
[Publications] S.Iwaoka: "Characterization of three cytochrome Pー450s purified from renal microsomes of untreated male rats and comparison with human renal cytochrome Pー450" Arch.Biochem.Biophys.276. 473-480 (1990)
[Publications] S.Iwaoka: "Induction and regulation of cytochrome P450Kー5(lauric acid hydroxylase) in rat renal microsomes by starvation" Biochim.Biophys.Acta. 1036. 8-23 (1990)
[Publications] Y.Sagara: "Structural organization of the bovine adrenodoxin gene" J.Biochem.107. 77-83 (1990)

1990 Fiscal Year Annual Research Report

チトクロ-ムPー450の分子生物学:発現の分子機構

Principal Investigator

大村 恒雄 九州大学, 大学院医学系研究科, 教授 (80029933)

Research Products

[Publications] T.Sato: "The Aminoーterminal Structures That Determine Topological Orientation of Cytochrome Pー450 in Microsomal Membrane" EMBO J.9. 2391-2397 (1990)

[Publications] Shinーichirou Honda: "Novel cAMP Regulatory Elements in the Promotor Region of Bovine Pー450(11β) Gene" J.Biochem.108. 1042-1049 (1990)

[Publications] Egawa,T: "Resonance Raman Study on Mutant Cytochrome Pー450 Obtained by SiteーDirected Mutagenesis" Biochim.Biophys.Acta. 1040. 211-216 (1990)

[Publications] Imai,Y: "Characterization of Rabbit Liver P450IIE1 Synthesized in Transformed Yeast Cells" J.Biochem.108. 522-524 (1990)

[Publications] M.Noshiro: "Rat Liver Cholesterol 7αーHydroxylase" J.Biol.Chem.265. 10036-10041 (1990)

[Publications] Yamazoe,Y: "Modulation of hepatic level of microsomal testosterone 7αーhydroxylase,Pー450a(Pー450IIA),by thyroid hormone and growth hormone in rat liver" J.Biochem.108. 599-603 (1990)

[Publications] Nagata,K: "Purification and characterization of four catalytically active testosterone 6βーhydroxylase Pー450s from rat liver microsomes: comparison of a novel form with three structurally and functionally related forms" J.Biochem.107. 718-725 (1990)

[Publications] M.Kitada: "Metabolic activation of aflatoxin B,and 2ーaminoー3ーmethylimidazo(4,5ーf)quinoline by human adult and fetal livers" Cancer Res.50. 2641-2645 (1990)

[Publications] T.Uchida: "Isolation of cDNA coding for three different forms of liver microsomal cytochrome Pー450 from polychlorinated biphenylーtreated beagle dogs" Mol.Pharmacol.38. 644-651 (1990)

[Publications] H.Sotokawa: "Electron Spin Resonance Studies of WildーType and Mutant Cytochromes Pー450_d: Effects of Mutations at Proximal,Aromatic and Distal Sites on g__ー Values" Biochim.Biophys.Acta. 1037. 122-128 (1990)

[Publications] Yamamoto,A: "Absence of cytochrome Pー450 and presence of autolysosomal membrane antigens on the isolation membranes and autophagosomal membranes in rat hepatocytes" J.Histochem.Cytochem.38. 1571-1581 (1990)

[Publications] Yokotani,N: "cDNA cloning of cytochrome Pー450 related to Pー450_pー2 from the expression" Eur.J.Biochem.187. 23-29 (1990)

[Publications] Yanagida,A: "A novel cisーacting DNA element required for a high level of inducible expression of the rat Pー450c gene" Mol.Cell.Biol.10. 1470-1475 (1990)

[Publications] S.Iwaoka: "Characterization of three cytochrome Pー450s purified from renal microsomes of untreated male rats and comparison with human renal cytochrome Pー450" Arch.Biochem.Biophys.276. 473-480 (1990)

[Publications] S.Iwaoka: "Induction and regulation of cytochrome P450Kー5(lauric acid hydroxylase) in rat renal microsomes by starvation" Biochim.Biophys.Acta. 1036. 8-23 (1990)

[Publications] Y.Sagara: "Structural organization of the bovine adrenodoxin gene" J.Biochem.107. 77-83 (1990)

大村恒雄九州大学, 大学院医学系研究科, 教授 (80029933)