1989 Fiscal Year Annual Research Report

チトクロームPー450の分子生物学:発現の分子機構

Research Project

Project/Area Number	01635001
Research Institution	Tohoku University
Principal Investigator	藤井義明東北大学, 理学部, 教授 (00098146)
Co-Investigator(Kenkyū-buntansha)	船江良彦大阪市立大学, 医学部, 助教授 (00047268) 田代裕関西医科大学, 第一生理学, 教授 (40077558) 清水透東北大学, 非水溶液化学研究所, 助教授 (40118956) 楠瀬正道大阪市立大学, 医学部附属刀根山結核研究所, 教授 (10046766) 川尻要埼玉県立がんセンター研究所, 生化学部, 主任研究員 (50142112)
Keywords	ヘム蛋白質 / 脂溶性生理活性物質 / 薬物の酸化的代謝 / 分子多様性 / 細胞内局在性 / ミトコンドリア / ミクロソーム / 遺伝子発現
Research Abstract	Pー450分子生物学、発現の分子機構の本年の研究実績は、1)分子多様性、基質特異性に関するもの、2)遺伝子発現制御に関するもの、3)細胞内局在性に関するものの3つに大別することができる。1)に関する研究の発展は、今井、清水、堀内によって既にクローン化されているPー450のcDNAを用いて部位特異的変異の導入あるいは、Pー450間の部分的構造の交換を行って、基質の結合部位、還元酵素と相互作用する部位などが明らかにされつつある。奥田は肝汁酸合成の律速酵素である7α水酸酵素とビタミンD_324水酸化酵素のPー450cDNAを分離してその塩基配列を決定し、蛋白質の一次構造を演繹した。楠瀬はプロスタグランジンや脂肪酸のω水酸化反応を触媒する三種類のPー450のcDNAの単離及びその構造決定を行い、アミノ酸配列を明らかにした。鎌滝PCBあるいは3MC処理のカニクイザル肝から、渡辺は3ーMC処理ハムスター肝から各々Pー450のcDNAクローンを単離し構造を決定した。川尻はPー450IAl遺伝子に多型の存在することを明らかにし、その一つの型と肺癌との関係を示唆する結果を得た。今後例数を増やして正否の確認が望まれる。2)については、加藤がPー450f、jあるいは雌雄特異的Pー450が成長ホルモンによって正、負の調節をうけていることを示した。大村はPー45011βの遺伝子のcAMPによる調節領域と副腎で特異的に発現する制御領域を欠失変異の導入とin vitroの転写系を用いて決定した。薬物によって制御を受けるPー450Cについて、藤井はXREとBTEの2種の制御領域によって発現制御されていることを明らかにしたが、BTEに結合する転写因子のcDNAクローニングを行い、その構造解析を行っている。船江はXREに働く転写因子の精製をXRE結合カラムなどを用いて続けている。3)について田代は改善した定量的免疫電顕法を用いて、フェノバルビタール投与後のPー450の分解過程でリソゾームの関与を明らかにした。

Research Products

(18 results)

All Other

All Publications (18 results)

[Publications] Yoshio Imai.: "Point Mutations at Threonine-301 Modify Substrate Specificity of Rabbit Liver Microsomal Cytochromes P-450(Laurate(ω-1)-Hydroxylase and Testosterone 16 α-Hydroxylase)" Biochem.Biophys.Res.Commun.158. 717-722 (1989)
[Publications] Tomohide Uno.: "Identification of Regions Functioning in Substrate Interaction of Rabbit Liver Cytochrome P-450(Laurate(ω-1)-Hydroxylase)" J.Biochem.106. 569-574 (1989)
[Publications] Furuya,H.: "Site-Directed Mutageneses of Rat Liver Cytochrome P-450d:Catalytic Activities toward Benzphetamine and 7-Ethoxycoumarin" Biochemistry. 28. 6848-6857 (1989)
[Publications] Furuya,H.: "Mutations at the Distal and Proximal Sites of Cytochrome P-450s Changed Regio-Specificity of Acetanilide Hydroxylations" Biochem.Biophys.Res.Commun.160. 669-676 (1989)
[Publications] Imai,M.: "Unicoupling of the cytochrome P-450cam monooxygenase reaction by a single mutation of threonine 252 to alanine or valin:A possible role of the hydroxy amino acid in the oxygen activation" Proc.Natl.Acad.Sci.U.S.A.86. 7823-7827 (1989)
[Publications] Okuda,K.: "Purification of 25-hydroxyvitamin D_3 24-hydroxylase from rat kidney mitochondria." FEBS Lett.255. 405-408 (1989)
[Publications] Okuda,K.: "Molecular cloning of cDNA for cholesterol 7 α-hydroxylase from rat liver microsomes:Nucleotide sequence and expression." FEBS Lett.257. 97-100 (1989)
[Publications] Y.Kikuta.: "Purification and characterization of hepatic microsomal prostaglandin ω-hydroxylase cytochrome P-450 from pregnant rabbits." J.Biochem.106. 468-473 (1989)
[Publications] N.Yokotani.: "Two forms of ω-hydroxylase toward prostaglandin A and laurate;cDNA cloning and their expression." J.Biol.Cham.264. 21665-21669 (1989)
[Publications] Masayuki Komori.: "Molecularcloning and sequence analysis of cDNA containing the entire coding region for human fetal liver cytochrome P-450." J.Biochem.105. 161-163 (1989)
[Publications] Masayuki Komori.: "Isolation of a new human fetal liver cytochrome P-450 cDNA clone:Evidence for expression of a limited number of form(s)fo cytochrome P-450 in human fetal livers." Archs.Biochem.Biophys.272. 219-255 (1989)
[Publications] Ohmachi,T.: "Purification and characterization of cytochrome P-450 from liver microsomes of 3-methylcholanthrene-treated crab-eating monkeys" J.Biochem.105. 307-311 (1989)
[Publications] Wong,G.: "Gene family of male-specific testosterone 16 α-hydroxylase(C-P-450_<16α>)in mice." J.Biol.Chem.264. 2920-2927 (1989)
[Publications] Shimada,M.: "Suppression in expression of a male-specific cytochrome P450,P450-male:Difference in the effect of chemical incucers on P450-male mRNA and protein in rat livers," Arch.Biochem.Biophys.270. 578-587 (1989)
[Publications] Yamazoe,Y.: "Cytochrome P-450 in livers of diabetic rats:Regulation by growth hormone and insulin" Arch.Biochem.Biophys.268. 567-575 (1989)
[Publications] Toshihide Hashimote.: "Cloning and Characterization of Bovine Cytochrome P-450(11β)Genes." J.Biochem.105. 676-679 (1989)
[Publications] Wei-Jia Ou.: "Purification and Characterization of a Processing Protease from Rat Liver Mitochondria." EMBO J.8. 2605-2612 (1989)
[Publications] Imaoka,S.: "Expression of four phenobarbital-inducible cytochrome P-450s in liver,kidney,and lung of rats." J.Biochem.105. 939-945 (1989)

1989 Fiscal Year Annual Research Report

チトクロームPー450の分子生物学:発現の分子機構

Principal Investigator

藤井 義明 東北大学, 理学部, 教授 (00098146)

Research Products

[Publications] Yoshio Imai.: "Point Mutations at Threonine-301 Modify Substrate Specificity of Rabbit Liver Microsomal Cytochromes P-450(Laurate(ω-1)-Hydroxylase and Testosterone 16 α-Hydroxylase)" Biochem.Biophys.Res.Commun.158. 717-722 (1989)

[Publications] Tomohide Uno.: "Identification of Regions Functioning in Substrate Interaction of Rabbit Liver Cytochrome P-450(Laurate(ω-1)-Hydroxylase)" J.Biochem.106. 569-574 (1989)

[Publications] Furuya,H.: "Site-Directed Mutageneses of Rat Liver Cytochrome P-450d:Catalytic Activities toward Benzphetamine and 7-Ethoxycoumarin" Biochemistry. 28. 6848-6857 (1989)

[Publications] Furuya,H.: "Mutations at the Distal and Proximal Sites of Cytochrome P-450s Changed Regio-Specificity of Acetanilide Hydroxylations" Biochem.Biophys.Res.Commun.160. 669-676 (1989)

[Publications] Imai,M.: "Unicoupling of the cytochrome P-450cam monooxygenase reaction by a single mutation of threonine 252 to alanine or valin:A possible role of the hydroxy amino acid in the oxygen activation" Proc.Natl.Acad.Sci.U.S.A.86. 7823-7827 (1989)

[Publications] Okuda,K.: "Purification of 25-hydroxyvitamin D_3 24-hydroxylase from rat kidney mitochondria." FEBS Lett.255. 405-408 (1989)

[Publications] Okuda,K.: "Molecular cloning of cDNA for cholesterol 7 α-hydroxylase from rat liver microsomes:Nucleotide sequence and expression." FEBS Lett.257. 97-100 (1989)

[Publications] Y.Kikuta.: "Purification and characterization of hepatic microsomal prostaglandin ω-hydroxylase cytochrome P-450 from pregnant rabbits." J.Biochem.106. 468-473 (1989)

[Publications] N.Yokotani.: "Two forms of ω-hydroxylase toward prostaglandin A and laurate;cDNA cloning and their expression." J.Biol.Cham.264. 21665-21669 (1989)

[Publications] Masayuki Komori.: "Molecularcloning and sequence analysis of cDNA containing the entire coding region for human fetal liver cytochrome P-450." J.Biochem.105. 161-163 (1989)

[Publications] Masayuki Komori.: "Isolation of a new human fetal liver cytochrome P-450 cDNA clone:Evidence for expression of a limited number of form(s)fo cytochrome P-450 in human fetal livers." Archs.Biochem.Biophys.272. 219-255 (1989)

[Publications] Ohmachi,T.: "Purification and characterization of cytochrome P-450 from liver microsomes of 3-methylcholanthrene-treated crab-eating monkeys" J.Biochem.105. 307-311 (1989)

[Publications] Wong,G.: "Gene family of male-specific testosterone 16 α-hydroxylase(C-P-450_<16α>)in mice." J.Biol.Chem.264. 2920-2927 (1989)

[Publications] Shimada,M.: "Suppression in expression of a male-specific cytochrome P450,P450-male:Difference in the effect of chemical incucers on P450-male mRNA and protein in rat livers," Arch.Biochem.Biophys.270. 578-587 (1989)

[Publications] Yamazoe,Y.: "Cytochrome P-450 in livers of diabetic rats:Regulation by growth hormone and insulin" Arch.Biochem.Biophys.268. 567-575 (1989)

[Publications] Toshihide Hashimote.: "Cloning and Characterization of Bovine Cytochrome P-450(11β)Genes." J.Biochem.105. 676-679 (1989)

[Publications] Wei-Jia Ou.: "Purification and Characterization of a Processing Protease from Rat Liver Mitochondria." EMBO J.8. 2605-2612 (1989)

[Publications] Imaoka,S.: "Expression of four phenobarbital-inducible cytochrome P-450s in liver,kidney,and lung of rats." J.Biochem.105. 939-945 (1989)

藤井義明東北大学, 理学部, 教授 (00098146)