1994 Fiscal Year Annual Research Report

微繋蛋白質の末端配列決定法、特に新しい力ルポキシ末端配列の決定法の確立

Research Project

Project/Area Number	04102008
Research Institution	SCIENCE UNIVERSITY OF TOKYO,RESEARCH INSTITUTE OF BIOSCIENCES
Principal Investigator	次田晧東京理科大学, 総合研究所, 教授 (00028284)
Co-Investigator(Kenkyū-buntansha)	加茂正晴東京理科大学, 総合研究所, 助手 (40214564)
Keywords	C末端酸配分折 / N末端配列分析 / 不均一相微量反応 / 二次元電気泳鋤 / 過フッ素酸無水物 / 質量分析
Research Abstract	1)酸無水物を用いるC末端アミノ酸配列決定法;バンタフロロプロビオン酸無水物、ヘブタフロロ酪酸無水物をべブチドに-20℃1時間反応させるとC末端から一つづつアミノ酸の外れた分解混合物を得る事を見出しだが、その際反応の再現性を空気中の湿気が阻害している事を実験的に見出し、全実験をグロープポックス中で乾燥窒素ガスを流し乍ら実験を行う事とした。又無水物のアセト二トリル溶液からの気相反応が水分の除去に貢献している事を見出した。この反応の機構の研究により中間生成物としてアルコールでヱステルを作るオキサゾロン又は混合酸無水物を見出した。又この反応における副生成物について主生成物より-1、-8-46の各質量イオンを主反応生成物の他に見出し、その各々を物質的に確認した。 2)N末端配列についてはエドマン分解の中間生成物のアミノ化には不均一相反応が微重反応に有効である事を見出し、その線にそったフミノ化高感度化の端緒を開いた。又ヒドラジンの脱ブロック反応を二次元電気泳勅で得られた60ヶのN末端ブロックの蛋白質に適用しその約1割のデプロツクに成功した。他のアセチルのデプロックの反応を見出しつつある。 3)二次元電気泳動ではイネの蛋白質に引き続き実験植物としてゲノム検索の始められているシロイヌナズナについて行い5組織の蛋白質について合計約5,000ヶの蛋白質を分離し、内約100ヶの蛋白質の配列分析を進め約60ヶの蛋白部分構造を得た。続いてフザリウム菌についても同様の研究を進め1,500ヶの蛋白を分離、その約8%の蛋白質の解析を行った。

Research Products
(21 results)

All Other

All Publications (21 results)

[Publications] H.Iwadate: "Amino acid segeunce of ferredoxin-thioredoxin reductass variable subunit" Eur.J.Biochem.223. 465-471 (1994)
[Publications] M.Kamo: "Two dimensional electrophoresis of rice proteins" Workshop on Rice MoI.BioI.57- (1994)
[Publications] M.Kamo: "Characterization of proteins from rice and Arabidopsis with two dimensional electrphoresis" Protein Engineering. 7. 1161-1162 (1994)
[Publications] M.Kamo: "Separation and characterization of proteins with two-dimensional electrophoresis" J.Prot.Chem.13. 475-476 (1994)
[Publications] M.Kamo: "Separation & characterization of plant proteins with two dimensional electrophoresis" rotein Science-. 31. 97- (1994)
[Publications] N.Miyatake: "A specific chemical cleavage of asparaginyl and glycyl-glycine bonds in peptides and proteins by avapornhydrous hydrazine" J.Biochem.115. 208-212 (1994)
[Publications] M.Miyazaki: "Identification as b-adducin of a protein interacting with raphilin-3A in the presence of Ca2+ and phosphatidglserine21GC07:Biochem.Biophys.Res.Commun." 205. 460-466 (1994)
[Publications] Y.Nabuchi: "C-terminal sequencing of peptides and proteins by successive degradation with heptafluorobutylic anhydride vapor" Chem.Lett.757-760 (1994)
[Publications] K.Takamoto: "An amino acid sequencing method using perfluoroacyl anhydride" Protein Engineering. 7. 1167 (1994)
[Publications] A.Tsugita: "Two dimensional electrophoresis of rice proteins and novel C-terminal sequencing methods" The 23rd Seminar on Science and Technology. 91-100 (1994)
[Publications] A.Tsugita: "Separation and characterization of plant proteins with two dimensional electrophoresis" Proc.Japan-ROC Symposium on Agricultural Biotech.3-5 (1994)
[Publications] A.Tsugita: "Separation and characterization of plant proteins with two dimensional electrophoresis" Workshop on Rice MoI.BioI. 12-13 (1994)
[Publications] A.Tsugita: "A novel C-terminal sequencing method using perfluoroacyl anhydrides" J.Protein Chem.13. 476-477 (1994)
[Publications] A.Tsugita: "C-terminal sequencing method using perfluoroacyl anhydrides" Protein Science. 13. 78 (1994)
[Publications] A.Tsugita: "Separation and characterization ofrice protein" Electrophoresis. 15. 708-720 (1994)
[Publications] L.P.Chow: "Two dimensional polyacrylamide gel electrophoresis of Fusarium sporotrichioides proteins" J.Biomed.Sci.(発表予定).
[Publications] K.Horimoto: "Protein superfamily database (PROSUP)" Proc.14th International CODATA Conference. (発表予定).
[Publications] K.Horimoto: "A method for detecting an obscure sequence motif:application to DNA binding motifs" Prot.Eng.(発表予定).
[Publications] M.Kamo: "Separation and characterization ofArabidopsis proteins by two-dimensional gel electrophoresis" Electrophoresis. (発表予定).
[Publications] K.Takamoto: "Carboxy-terminal degradation of peptides using perfluoroacyl anhydrides a novel C-terminal seguencing method" Eur.J.Biochem.(発表予定).
[Publications] A.Tsugita: "A C-terminal seguencing method using perfluoroacyl anhydrides vapor method in protein structure analysis" Plenum Pub.(発表予定).

1994 Fiscal Year Annual Research Report

微繋蛋白質の末端配列決定法、特に新しい力ルポキシ末端配列の決定法の確立

Principal Investigator

次田 晧 東京理科大学, 総合研究所, 教授 (00028284)

Research Products

[Publications] H.Iwadate: "Amino acid segeunce of ferredoxin-thioredoxin reductass variable subunit" Eur.J.Biochem.223. 465-471 (1994)

[Publications] M.Kamo: "Two dimensional electrophoresis of rice proteins" Workshop on Rice MoI.BioI.57- (1994)

[Publications] M.Kamo: "Characterization of proteins from rice and Arabidopsis with two dimensional electrphoresis" Protein Engineering. 7. 1161-1162 (1994)

[Publications] M.Kamo: "Separation and characterization of proteins with two-dimensional electrophoresis" J.Prot.Chem.13. 475-476 (1994)

[Publications] M.Kamo: "Separation & characterization of plant proteins with two dimensional electrophoresis" rotein Science-. 31. 97- (1994)

[Publications] N.Miyatake: "A specific chemical cleavage of asparaginyl and glycyl-glycine bonds in peptides and proteins by avapornhydrous hydrazine" J.Biochem.115. 208-212 (1994)

[Publications] M.Miyazaki: "Identification as b-adducin of a protein interacting with raphilin-3A in the presence of Ca2+ and phosphatidglserine21GC07:Biochem.Biophys.Res.Commun." 205. 460-466 (1994)

[Publications] Y.Nabuchi: "C-terminal sequencing of peptides and proteins by successive degradation with heptafluorobutylic anhydride vapor" Chem.Lett.757-760 (1994)

[Publications] K.Takamoto: "An amino acid sequencing method using perfluoroacyl anhydride" Protein Engineering. 7. 1167 (1994)

[Publications] A.Tsugita: "Two dimensional electrophoresis of rice proteins and novel C-terminal sequencing methods" The 23rd Seminar on Science and Technology. 91-100 (1994)

[Publications] A.Tsugita: "Separation and characterization of plant proteins with two dimensional electrophoresis" Proc.Japan-ROC Symposium on Agricultural Biotech.3-5 (1994)

[Publications] A.Tsugita: "Separation and characterization of plant proteins with two dimensional electrophoresis" Workshop on Rice MoI.BioI. 12-13 (1994)

[Publications] A.Tsugita: "A novel C-terminal sequencing method using perfluoroacyl anhydrides" J.Protein Chem.13. 476-477 (1994)

[Publications] A.Tsugita: "C-terminal sequencing method using perfluoroacyl anhydrides" Protein Science. 13. 78 (1994)

[Publications] A.Tsugita: "Separation and characterization ofrice protein" Electrophoresis. 15. 708-720 (1994)

[Publications] L.P.Chow: "Two dimensional polyacrylamide gel electrophoresis of Fusarium sporotrichioides proteins" J.Biomed.Sci.(発表予定).

[Publications] K.Horimoto: "Protein superfamily database (PROSUP)" Proc.14th International CODATA Conference. (発表予定).

[Publications] K.Horimoto: "A method for detecting an obscure sequence motif:application to DNA binding motifs" Prot.Eng.(発表予定).

[Publications] M.Kamo: "Separation and characterization ofArabidopsis proteins by two-dimensional gel electrophoresis" Electrophoresis. (発表予定).

[Publications] K.Takamoto: "Carboxy-terminal degradation of peptides using perfluoroacyl anhydrides a novel C-terminal seguencing method" Eur.J.Biochem.(発表予定).

[Publications] A.Tsugita: "A C-terminal seguencing method using perfluoroacyl anhydrides vapor method in protein structure analysis" Plenum Pub.(発表予定).

次田晧東京理科大学, 総合研究所, 教授 (00028284)