1997 Fiscal Year Annual Research Report

エンド型グリコシダーゼの特異的な活性を利用した新しい糖鎖工学的研究の展開

Research Project

Project/Area Number	09358013
Research Category	Grant-in-Aid for Scientific Research (A)
Research Institution	Hirosaki University
Principal Investigator	遠藤正彦弘前大学, 医学部, 教授 (20004616)
Co-Investigator(Kenkyū-buntansha)	伊藤和央大阪市立大, 理学部, 講師 (20183171) 竹川薫香川大学, 農学部, 助教授 (50197282) 伊東信九州大学, 農学部, 助教授 (40253512) 山本憲二京都大学, 農学部, 助教授 (70109049) 堀田恭子北里大学, 医学部, 教授 (10050402)
Keywords	エンド型グリコシダーゼ / 糖鎖工学 / 糖タンパク質 / 糖脂質 / プロテオグリカン / 糖鎖切り出し / 糖鎖組み換え / 糖鎖限定分解
Research Abstract	目的と背景:本研究組織構成員はそれぞれがエンド型グリコシダーゼを発見した。その酵素は、複合糖質のコア部分(タンパク質または脂質)と糖鎖の結合部位(橋渡し部位)を選択的に分解するという特異的な性質を有する。したがって、本酵素は糖鎖の切り出し、限定分解、及びこの加水分解反応の逆反応としての糖転移反応を利用すると、新しい糖鎖工学的研究の展開が期待される。本研究は、本酵素群の分離法の確立と新しい糖鎖工学的研究展開を図ることにある。研究の実績:当研究構成員は、以下のようなエンド型グリコシダーゼを発見した。 1.プロテオグリカンに作用するエンド-β-キシロシダーゼ、エンド-β-ガラクトシダーゼ、エンド-β-グルクロニダーゼ(遠藤・高垣) 2.O-グリコシド型糖タンパク質に作用するエンド-α-α-N-アセチルガラクトサミニダーゼ(堀田) 3.N-グリコシド型糖タンパク質に作用するエンド-β-N-アセチルグルコサミニダーゼ(山本、竹川、伊藤) 4.糖脂質に作用するエンドグリコセラミダーゼ(伊東) これらの酵素について、(1)活性測定法の確立、(2)精製法の確立、(3)酵素の基質特異性について検討が進められてきた。今後の課題:酵素の基質特異性が明らかになった段階で、各種の糖タンパク質へ応用し、限定分解への適用の確立を図ることと、糖鎖の組み換えに向けて応用的展開の準備を進める予定である。

Research Products
(15 results)

All Other

All Publications (15 results)

[Publications] Takagaki,K.et al.: "Regulation of hyaluronate metabolism by progesterone in cultured skin fibroblasts from the human uterine cervix." FEBS Letters. 402. 223-226 (1997)
[Publications] Tanaka,K.et al.: "Effect of monensin on the synthesis of β-D-xyloside-induced glycosaminoglycan and its likage region oligosaccharides in human skin fibroblasts." Journal of Biochemistry. 122. 1129-1132 (1997)
[Publications] Yasukochi,T.et al.: "Enzymatic synthesis of 4-methylubelliferyl glycosides of trisaccharide,and core tetrasaccharide,Gal(β1-3)Gal(β1-4)Xyl and GlcA(β1-3)Gal(β1-3)Gal(β1-4)Xvl,corresponding to the linkage region of proteoglycans." Bull.Chem.Soc.Jpn.70. 2719-2725 (1997)
[Publications] Takagaki,K.et al.: "Ion-spray mass spectromentry for identification of the nonreducing terminal sugar of glycosaminoglycan." Glycobiology. 8(印刷中). (1998)
[Publications] Takagaki,K.et al.: "Characterization of β-D-xyloside-initiated glycosaminoglycan synthesized by human skin fibroblasts in the presence of tunicamycin." Glycoconjugate Jounal. 15(印刷中). (1998)
[Publications] Tazawa,T.et al: "A novel 4-methylumbellifery1-β-D-xyloside derivative,sulfate-0-3-xylosylβ1-(4-methylumbelliferone),isolated from culture medium of human skin fibroblasts,and its role in methylumbelliferone-initiated glycosaminoglycan biosynthesis." Glycobiology. 8(印刷中). (1998)
[Publications] Kadowaki,S.et al.: "Transglycosylation activity of β-N-acetylhexosaminidase from Penicillium oxalicum and its application to synthesis of a drug carrier." J.Ferment.Bioeng.83(4). 341-345 (1997)
[Publications] Yamamoto,K. and Takegawa,K.: "Transglycosylation activity of microbial endoglycosidases and its application." Trends Glycosci.Glycotechnol.9. 339-354 (1997)
[Publications] Yamamoto,K.et al.: "Chemo-enzymatic synthesis of a novel glycopeptide using a microbial endoglycosidase." Carbohyd.Res.305(輸入中のため不明). (1997)
[Publications] 竹川薫: "エンドグリコシダーゼの糖転移反応を利用したネオ複合糖質の合成." バイオサイエンスとインダストリー. 20-26 (1997)
[Publications] Takegawa,K.et al.: "Enzymatic synthesis of a neoglycoconjigate by transglycosylation with arthrobacter endo-b-N-acetylglucosaminidase:A substrate for colorimetric detection of endo-b-N-acetylglucosaminidase activity." Anal.Biochem.(印刷中). (1998)
[Publications] Ishii-Karakasa,I.et al.: "Structual determination of the 0-linked sialyl oligosaccharides liberated from fetuin with endo-α-N-acetylgalactosaminidase-S by HLPC analysis and 500-MHz ^1H-NMR spectroscopy." Eur.J.Biochem.247. 709-715 (1997)
[Publications] Mitsutake,S.et al.: "Synthesis of ^<14>C-labeled ceramide by fatty acid trasfer reaction of sphigolipid ceramideN-deacylase and its application for assay of ceramidase." Analyt.biochem.247. 52-57 (1997)
[Publications] Izu,Y.et al.: "Molecular cloning,expression,and sequence analysis of the endoglycoceramidase II gene from Rhodocococcus sp.strain M-777." J.Biol.Chem.272. 19846-19850 (1997)
[Publications] Mistutake,S.et al.: "Enzymatic synthesis of [^<14>C]-glycosphingolipids by reverse hydrolysis reaction of sphingolipid ceramide N-deacylase:Application for detection of EGCase activity." J.Biochem.(印刷中). (1998)

1997 Fiscal Year Annual Research Report

エンド型グリコシダーゼの特異的な活性を利用した新しい糖鎖工学的研究の展開

Principal Investigator

遠藤 正彦 弘前大学, 医学部, 教授 (20004616)

Research Products

[Publications] Takagaki,K.et al.: "Regulation of hyaluronate metabolism by progesterone in cultured skin fibroblasts from the human uterine cervix." FEBS Letters. 402. 223-226 (1997)

[Publications] Tanaka,K.et al.: "Effect of monensin on the synthesis of β-D-xyloside-induced glycosaminoglycan and its likage region oligosaccharides in human skin fibroblasts." Journal of Biochemistry. 122. 1129-1132 (1997)

[Publications] Yasukochi,T.et al.: "Enzymatic synthesis of 4-methylubelliferyl glycosides of trisaccharide,and core tetrasaccharide,Gal(β1-3)Gal(β1-4)Xyl and GlcA(β1-3)Gal(β1-3)Gal(β1-4)Xvl,corresponding to the linkage region of proteoglycans." Bull.Chem.Soc.Jpn.70. 2719-2725 (1997)

[Publications] Takagaki,K.et al.: "Ion-spray mass spectromentry for identification of the nonreducing terminal sugar of glycosaminoglycan." Glycobiology. 8(印刷中). (1998)

[Publications] Takagaki,K.et al.: "Characterization of β-D-xyloside-initiated glycosaminoglycan synthesized by human skin fibroblasts in the presence of tunicamycin." Glycoconjugate Jounal. 15(印刷中). (1998)

[Publications] Kadowaki,S.et al.: "Transglycosylation activity of β-N-acetylhexosaminidase from Penicillium oxalicum and its application to synthesis of a drug carrier." J.Ferment.Bioeng.83(4). 341-345 (1997)

[Publications] Yamamoto,K. and Takegawa,K.: "Transglycosylation activity of microbial endoglycosidases and its application." Trends Glycosci.Glycotechnol.9. 339-354 (1997)

[Publications] Yamamoto,K.et al.: "Chemo-enzymatic synthesis of a novel glycopeptide using a microbial endoglycosidase." Carbohyd.Res.305(輸入中のため不明). (1997)

[Publications] 竹川 薫: "エンドグリコシダーゼの糖転移反応を利用したネオ複合糖質の合成." バイオサイエンスとインダストリー. 20-26 (1997)

[Publications] Takegawa,K.et al.: "Enzymatic synthesis of a neoglycoconjigate by transglycosylation with arthrobacter endo-b-N-acetylglucosaminidase:A substrate for colorimetric detection of endo-b-N-acetylglucosaminidase activity." Anal.Biochem.(印刷中). (1998)

[Publications] Ishii-Karakasa,I.et al.: "Structual determination of the 0-linked sialyl oligosaccharides liberated from fetuin with endo-α-N-acetylgalactosaminidase-S by HLPC analysis and 500-MHz ^1H-NMR spectroscopy." Eur.J.Biochem.247. 709-715 (1997)

[Publications] Mitsutake,S.et al.: "Synthesis of ^<14>C-labeled ceramide by fatty acid trasfer reaction of sphigolipid ceramideN-deacylase and its application for assay of ceramidase." Analyt.biochem.247. 52-57 (1997)

[Publications] Izu,Y.et al.: "Molecular cloning,expression,and sequence analysis of the endoglycoceramidase II gene from Rhodocococcus sp.strain M-777." J.Biol.Chem.272. 19846-19850 (1997)

[Publications] Mistutake,S.et al.: "Enzymatic synthesis of [^<14>C]-glycosphingolipids by reverse hydrolysis reaction of sphingolipid ceramide N-deacylase:Application for detection of EGCase activity." J.Biochem.(印刷中). (1998)

遠藤正彦弘前大学, 医学部, 教授 (20004616)

[Publications] 竹川薫: "エンドグリコシダーゼの糖転移反応を利用したネオ複合糖質の合成." バイオサイエンスとインダストリー. 20-26 (1997)