1990 Fiscal Year Annual Research Report

基質と酵素間の精密構造認識と反応機構

Research Project

Project/Area Number	02250101
Research Institution	Tohoku University
Principal Investigator	小倉協三東北大学, 非水溶液化学研究所, 教授 (80006303)
Co-Investigator(Kenkyū-buntansha)	左右田健次東京大学, 化学研究所, 教授 (30027023) 中嶋暉躬東京大学, 薬学部, 教授 (50012597) 三川潮東京大学, 薬学部, 教授 (60012613)
Keywords	イソプレノイド / 生合成 / プレニルトランスフェラ-ゼ / Pー450 / イソフラボノイド / アストパラン / Dーアミノ酸 / トランスアミナ-ゼ
Research Abstract	小倉らはイソプレノイド生合成の炭素鎖延長反応を触媒するプレニルトランスフェラ-ゼのうち、C_<40>,C_<45>およびC_<50>までのE型鎖延長をそれぞれ触媒する3つの酵素には蛋白性の活性化因子が存在することを見いだし、その活性化機構を明らかにした。また、この活性化因子の発見により、これらの長鎖プレニル二リン酸合成酵素の高度精製が初めて可能となった。電気泳動的に均一になったソラネシル二リン酸合成酵素は34KDaのサブユニットからなるホモダイマ-蛋白質であり、それ自体の触媒活性は低いが、活性化因子の添加により著しく活性が増大すること、および、この活性化因子は、非水溶性の反応生成物を酵素の活性部位から取り除き、触媒のタ-ンオ-バ-を維持するように働くものであることを明らかにした。また、長鎖プレニル二リン酸合成酵素による反応生成物の鎖長を決定する要因を明らかにし、鎖延長反応をin vitroで制御できることを示した。三川らはイソフラボノイド生合成におけるPー450 関与の反応でフェニル基が分子内で転位する反応機序を証明し、類似の機構で他の多くの天然物生合成の反応を説明できることを示した。中島らはハチ毒ペプチドであるマストラパランの構造類似体を用いてカルモジュリン依存性の酵素の活性阻害効果を調べ、それを指標にして酵素の分子認識機構が追求できることを示した。左右田らは耐熱性細菌由来のDーアミノ酸のトランスアミナ-ゼの一次構造決定に続いて、活性中心のリジンをアルギニンまたはアラニンに変えた変異体を作り、前者が低速度ではあるが、アミノ基転移の触媒活性を示すのに対し、後者はそれ自身では触媒機能をもたないが、アルキルアミンの存在下では機能を発現することを見いだした。また、耐熱性Lーアスパラギン酸トランスアミナ-ゼの活性中心のリジンをSー(βーアミノエチル)システインに変えたものは野生型酵素の14%の活性を示すことがわかった。

Research Products
(17 results)

All Other

All Publications (17 results)

[Publications] 小倉,協三他: "Negative Ion Fast Atom BombardmentーTandem Mass Spectroーmetry for Structural Analysis of Isoprenoid Diphosphates" Analitical Biochemistry. 185. 182-186 (1990)
[Publications] 小倉,協三他: "Prenyltransferase Reaction Involving Enantiomeric Discrimination" Tetrahedron Letters. 31. 4189-4190 (1990)
[Publications] 小倉,協三他: "Enzymatic Synthesis of Glycinoprenols." Tehrahedron Letters. 32. 241-242 (1991)
[Publications] 小倉,協三他: "Formation of Z___ー,E___ー,E___ーーGeranylgeranyl Diphosphate by Rat Liver Microsomes" The Journal of Biological Chemistry. 266. 3458-3463 (1991)
[Publications] 小倉,協三他: "Variable Product Specificity of Microsomal Dehydrodolichyl Diphosphate Synthase from Rat Liver" The Journal of Biological Chemistry. 266. 3464-3468 (1991)
[Publications] 三川,潮他: "Reaction mechanism of oxidative rearrangement of flavanone in isoflavone biosynthesis" FEBS Letters. 12. 219-222 (1990)
[Publications] 三川,潮他: "Stable ketoーisomer of tetrahydroxynaphthalene in alkali:Theoretical support to the postーaromatic reduction in the biosynthesis of aromatic polyketides" Tetrahedron Letters. 31. 5905-5908 (1990)
[Publications] 三川,潮他: "Hydroxyacetosyringe in the major virulence gene activating factors in belldonna hairy root cultures,and isositol enhances its activity" Chemical and Pharmaceutical Bulletin. 38. 2063-2065 (1990)
[Publications] 三川,潮他: "Nonーstereospecific incorporation of (1,2ー^<13>C_2)ーacetate into tropane alkal〓ids in <Hyoscyamus>___ー <albus>___ー" Chemical and Pharmaceutical Bulletin. 38. 2066-2068 (1990)
[Publications] 三川,潮他: "Isofalvone synthase from cell suspension cultures of <Pueralia>___ー <lobata>___ー" Chemical and Pharmaceutical Bulletin. 38. 1942-1945 (1990)
[Publications] 三川,潮他: "Oneーstep conversion of flavanones into isoflavones:A new facile biomimetic synthesis of isoflavones" Tetrahedron Letters. 31. 7355-7356 (1990)
[Publications] 中嶋,暉躬他: "Interaction of Wasp Venom Peptide Mastoparan with calmodulin" Peptide Chemistry. 109-114 (1990)
[Publications] 中嶋,暉躬他: "A ^1HーNMR Study of the Solution Conformation of Icaria Chemotactic Paptide and its [Lys^7]Analog" Biochemical Biophysical Research Communications. 168. 596-603 (1990)
[Publications] 中嶋,暉躬他: "Role of Lysine Residue at 7th Position of Wasp Chemotactic Peptides" Biochemical Biophysical Research Communications. 168. 844-849 (1990)
[Publications] 中嶋,暉躬他: "Chemotactic Peptide from Ropalidian Wasp as Well as the Authentic Chemotactic Tripeptide Stimulates Two Distinct Pathways in Neutrophils,but the [Lysー7]Analog does Only One of Them" Biochemical Biophysical Research Communications. (1991)
[Publications] 左右田,健次他: "Substitution of Sー(βーAminoethyl)ーCysteine for Active Site Lysine of Thermostable Aspartate Aminotransferase:" Journal of Biochemistry. 108. 699-700 (1990)
[Publications] 左右田,健次他: "Effect of Substitution of Pyridoxal PhosphateーBinding Lysyl Residue of Thermostable DーAmino Acid Aminotransーferase by Arginine and Alanine" Biochemistry. 30. (1991)

1990 Fiscal Year Annual Research Report

基質と酵素間の精密構造認識と反応機構

Principal Investigator

小倉協三東北大学, 非水溶液化学研究所, 教授 (80006303)

Research Products

[Publications] 小倉,協三他: "Negative Ion Fast Atom BombardmentーTandem Mass Spectroーmetry for Structural Analysis of Isoprenoid Diphosphates" Analitical Biochemistry. 185. 182-186 (1990)

[Publications] 小倉,協三他: "Prenyltransferase Reaction Involving Enantiomeric Discrimination" Tetrahedron Letters. 31. 4189-4190 (1990)

[Publications] 小倉,協三他: "Enzymatic Synthesis of Glycinoprenols." Tehrahedron Letters. 32. 241-242 (1991)

[Publications] 小倉,協三他: "Formation of Z_ー,E_ー,E___ーーGeranylgeranyl Diphosphate by Rat Liver Microsomes" The Journal of Biological Chemistry. 266. 3458-3463 (1991)

[Publications] 小倉,協三他: "Variable Product Specificity of Microsomal Dehydrodolichyl Diphosphate Synthase from Rat Liver" The Journal of Biological Chemistry. 266. 3464-3468 (1991)

[Publications] 三川,潮他: "Reaction mechanism of oxidative rearrangement of flavanone in isoflavone biosynthesis" FEBS Letters. 12. 219-222 (1990)

[Publications] 三川,潮他: "Stable ketoーisomer of tetrahydroxynaphthalene in alkali:Theoretical support to the postーaromatic reduction in the biosynthesis of aromatic polyketides" Tetrahedron Letters. 31. 5905-5908 (1990)

[Publications] 三川,潮他: "Hydroxyacetosyringe in the major virulence gene activating factors in belldonna hairy root cultures,and isositol enhances its activity" Chemical and Pharmaceutical Bulletin. 38. 2063-2065 (1990)

[Publications] 三川,潮他: "Nonーstereospecific incorporation of (1,2ー^<13>C_2)ーacetate into tropane alkal〓ids in <Hyoscyamus>_ー <albus>_ー" Chemical and Pharmaceutical Bulletin. 38. 2066-2068 (1990)

[Publications] 三川,潮他: "Isofalvone synthase from cell suspension cultures of <Pueralia>_ー <lobata>_ー" Chemical and Pharmaceutical Bulletin. 38. 1942-1945 (1990)

[Publications] 三川,潮他: "Oneーstep conversion of flavanones into isoflavones:A new facile biomimetic synthesis of isoflavones" Tetrahedron Letters. 31. 7355-7356 (1990)

[Publications] 中嶋,暉躬他: "Interaction of Wasp Venom Peptide Mastoparan with calmodulin" Peptide Chemistry. 109-114 (1990)

[Publications] 中嶋,暉躬他: "A ^1HーNMR Study of the Solution Conformation of Icaria Chemotactic Paptide and its [Lys^7]Analog" Biochemical Biophysical Research Communications. 168. 596-603 (1990)

[Publications] 中嶋,暉躬他: "Role of Lysine Residue at 7th Position of Wasp Chemotactic Peptides" Biochemical Biophysical Research Communications. 168. 844-849 (1990)

[Publications] 中嶋,暉躬他: "Chemotactic Peptide from Ropalidian Wasp as Well as the Authentic Chemotactic Tripeptide Stimulates Two Distinct Pathways in Neutrophils,but the [Lysー7]Analog does Only One of Them" Biochemical Biophysical Research Communications. (1991)

[Publications] 左右田,健次他: "Substitution of Sー(βーAminoethyl)ーCysteine for Active Site Lysine of Thermostable Aspartate Aminotransferase:" Journal of Biochemistry. 108. 699-700 (1990)

[Publications] 左右田,健次他: "Effect of Substitution of Pyridoxal PhosphateーBinding Lysyl Residue of Thermostable DーAmino Acid Aminotransーferase by Arginine and Alanine" Biochemistry. 30. (1991)

1990 Fiscal Year Annual Research Report

基質と酵素間の精密構造認識と反応機構

Principal Investigator

小倉 協三 東北大学, 非水溶液化学研究所, 教授 (80006303)

Research Products

[Publications] 小倉,協三 他: "Negative Ion Fast Atom BombardmentーTandem Mass Spectroーmetry for Structural Analysis of Isoprenoid Diphosphates" Analitical Biochemistry. 185. 182-186 (1990)

[Publications] 小倉,協三 他: "Prenyltransferase Reaction Involving Enantiomeric Discrimination" Tetrahedron Letters. 31. 4189-4190 (1990)

[Publications] 小倉,協三 他: "Enzymatic Synthesis of Glycinoprenols." Tehrahedron Letters. 32. 241-242 (1991)

[Publications] 小倉,協三 他: "Formation of Z___ー,E___ー,E___ーーGeranylgeranyl Diphosphate by Rat Liver Microsomes" The Journal of Biological Chemistry. 266. 3458-3463 (1991)

[Publications] 小倉,協三 他: "Variable Product Specificity of Microsomal Dehydrodolichyl Diphosphate Synthase from Rat Liver" The Journal of Biological Chemistry. 266. 3464-3468 (1991)

[Publications] 三川,潮 他: "Reaction mechanism of oxidative rearrangement of flavanone in isoflavone biosynthesis" FEBS Letters. 12. 219-222 (1990)

[Publications] 三川,潮 他: "Stable ketoーisomer of tetrahydroxynaphthalene in alkali:Theoretical support to the postーaromatic reduction in the biosynthesis of aromatic polyketides" Tetrahedron Letters. 31. 5905-5908 (1990)

[Publications] 三川,潮 他: "Hydroxyacetosyringe in the major virulence gene activating factors in belldonna hairy root cultures,and isositol enhances its activity" Chemical and Pharmaceutical Bulletin. 38. 2063-2065 (1990)

[Publications] 三川,潮 他: "Nonーstereospecific incorporation of (1,2ー^<13>C_2)ーacetate into tropane alkal〓ids in <Hyoscyamus>___ー <albus>___ー" Chemical and Pharmaceutical Bulletin. 38. 2066-2068 (1990)

[Publications] 三川,潮 他: "Isofalvone synthase from cell suspension cultures of <Pueralia>___ー <lobata>___ー" Chemical and Pharmaceutical Bulletin. 38. 1942-1945 (1990)

[Publications] 三川,潮 他: "Oneーstep conversion of flavanones into isoflavones:A new facile biomimetic synthesis of isoflavones" Tetrahedron Letters. 31. 7355-7356 (1990)

[Publications] 中嶋,暉躬 他: "Interaction of Wasp Venom Peptide Mastoparan with calmodulin" Peptide Chemistry. 109-114 (1990)

[Publications] 中嶋,暉躬 他: "A ^1HーNMR Study of the Solution Conformation of Icaria Chemotactic Paptide and its [Lys^7]Analog" Biochemical Biophysical Research Communications. 168. 596-603 (1990)

[Publications] 中嶋,暉躬 他: "Role of Lysine Residue at 7th Position of Wasp Chemotactic Peptides" Biochemical Biophysical Research Communications. 168. 844-849 (1990)

[Publications] 中嶋,暉躬 他: "Chemotactic Peptide from Ropalidian Wasp as Well as the Authentic Chemotactic Tripeptide Stimulates Two Distinct Pathways in Neutrophils,but the [Lysー7]Analog does Only One of Them" Biochemical Biophysical Research Communications. (1991)

[Publications] 左右田,健次 他: "Substitution of Sー(βーAminoethyl)ーCysteine for Active Site Lysine of Thermostable Aspartate Aminotransferase:" Journal of Biochemistry. 108. 699-700 (1990)

[Publications] 左右田,健次 他: "Effect of Substitution of Pyridoxal PhosphateーBinding Lysyl Residue of Thermostable DーAmino Acid Aminotransーferase by Arginine and Alanine" Biochemistry. 30. (1991)

小倉協三東北大学, 非水溶液化学研究所, 教授 (80006303)

[Publications] 小倉,協三他: "Negative Ion Fast Atom BombardmentーTandem Mass Spectroーmetry for Structural Analysis of Isoprenoid Diphosphates" Analitical Biochemistry. 185. 182-186 (1990)

[Publications] 小倉,協三他: "Prenyltransferase Reaction Involving Enantiomeric Discrimination" Tetrahedron Letters. 31. 4189-4190 (1990)

[Publications] 小倉,協三他: "Enzymatic Synthesis of Glycinoprenols." Tehrahedron Letters. 32. 241-242 (1991)

[Publications] 小倉,協三他: "Formation of Z_ー,E_ー,E___ーーGeranylgeranyl Diphosphate by Rat Liver Microsomes" The Journal of Biological Chemistry. 266. 3458-3463 (1991)

[Publications] 小倉,協三他: "Variable Product Specificity of Microsomal Dehydrodolichyl Diphosphate Synthase from Rat Liver" The Journal of Biological Chemistry. 266. 3464-3468 (1991)

[Publications] 三川,潮他: "Reaction mechanism of oxidative rearrangement of flavanone in isoflavone biosynthesis" FEBS Letters. 12. 219-222 (1990)

[Publications] 三川,潮他: "Stable ketoーisomer of tetrahydroxynaphthalene in alkali:Theoretical support to the postーaromatic reduction in the biosynthesis of aromatic polyketides" Tetrahedron Letters. 31. 5905-5908 (1990)

[Publications] 三川,潮他: "Hydroxyacetosyringe in the major virulence gene activating factors in belldonna hairy root cultures,and isositol enhances its activity" Chemical and Pharmaceutical Bulletin. 38. 2063-2065 (1990)

[Publications] 三川,潮他: "Nonーstereospecific incorporation of (1,2ー^<13>C_2)ーacetate into tropane alkal〓ids in <Hyoscyamus>_ー <albus>_ー" Chemical and Pharmaceutical Bulletin. 38. 2066-2068 (1990)

[Publications] 三川,潮他: "Isofalvone synthase from cell suspension cultures of <Pueralia>_ー <lobata>_ー" Chemical and Pharmaceutical Bulletin. 38. 1942-1945 (1990)

[Publications] 三川,潮他: "Oneーstep conversion of flavanones into isoflavones:A new facile biomimetic synthesis of isoflavones" Tetrahedron Letters. 31. 7355-7356 (1990)

[Publications] 中嶋,暉躬他: "Interaction of Wasp Venom Peptide Mastoparan with calmodulin" Peptide Chemistry. 109-114 (1990)

[Publications] 中嶋,暉躬他: "A ^1HーNMR Study of the Solution Conformation of Icaria Chemotactic Paptide and its [Lys^7]Analog" Biochemical Biophysical Research Communications. 168. 596-603 (1990)

[Publications] 中嶋,暉躬他: "Role of Lysine Residue at 7th Position of Wasp Chemotactic Peptides" Biochemical Biophysical Research Communications. 168. 844-849 (1990)

[Publications] 中嶋,暉躬他: "Chemotactic Peptide from Ropalidian Wasp as Well as the Authentic Chemotactic Tripeptide Stimulates Two Distinct Pathways in Neutrophils,but the [Lysー7]Analog does Only One of Them" Biochemical Biophysical Research Communications. (1991)

[Publications] 左右田,健次他: "Substitution of Sー(βーAminoethyl)ーCysteine for Active Site Lysine of Thermostable Aspartate Aminotransferase:" Journal of Biochemistry. 108. 699-700 (1990)

[Publications] 左右田,健次他: "Effect of Substitution of Pyridoxal PhosphateーBinding Lysyl Residue of Thermostable DーAmino Acid Aminotransーferase by Arginine and Alanine" Biochemistry. 30. (1991)