Identification of endo-type chondroitin sulfate hydrolases applicable to neuronal regenerative therapy

适用于神经元再生治疗的内型硫酸软骨素水解酶的鉴定

• 批准号: 21590057
• 负责人: YAMADA Shuhei
• 金额: $ 2.91万
• 依托单位: Hokkaido University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2009
• 资助国家: 日本
• 起止时间: 2009 至 2011
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-21590057/
• 关键词: グリコサミノグリカン; コンドロイチン硫酸; ヒアルロン酸; 神経再生; ヒアル; ヒアルロニダーゼ

In this study, human hyaluronidase-4 (HYAL4) was demonstrated to be a chondroitin sulfate (CS)-specific endo-type hydrolase for the first time. Although mouse hyaluronidase-4 (mHyal4) was also demonstrated to be a CS-specific endo-type hydrolase, it has broader substrate specificity than HYAL4. A series of human/mouse hyaluronidase-4 chimeric proteins and point mutants were generated, and the amino acid residues responsible for the difference in substrate specificity were identified. To prepare an active enzyme applicable to neuronal regenerative therapy in a large scale, the expression system in yeast was constructed. Injury region was extracted from the rats with lateral hemisection of the spinal cord, and the composition of CS in the region as well as its sensitivity to mHyal4 was investigated.

本研究首次证实人透明质酸酶-4（HYAL-4）是一种硫酸软骨素（CS）特异性内切型水解酶。尽管小鼠透明质酸酶-4（mHyal 4）也被证明是CS特异性内切型水解酶，但它比HYAL 4具有更广泛的底物特异性。产生了一系列人/小鼠透明质酸酶-4嵌合蛋白和点突变体，并鉴定了导致底物特异性差异的氨基酸残基。为了大规模制备适用于神经再生治疗的活性酶，构建了酵母表达系统。从脊髓半横断损伤大鼠中提取损伤区，研究损伤区CS的组成及其对mHyal 4的敏感性。

项目成果

マウスにおけるコンドロイチン硫酸特異的加水分解酵素の同定

小鼠硫酸软骨素特异性水解酶的鉴定

• 发表时间: 2010
• 作者: 田渕明子;袴田知之;石川充;塩田惇;庄司しずく;福地守;津田正明;宮崎杏
• 通讯作者: 宮崎杏

Structural and functional changes of sulfated glycosaminoglycans in Xenopus laevis during embryogenesis.

非洲爪蟾胚胎发生过程中硫酸化糖胺聚糖的结构和功能变化。

• 发表时间: 2009
• 期刊: Glycobiology 19(5)
• 作者: Yamada S;Onishi M;Fujinawa R;Tadokoro Y;Okabayashi K;Asashima M;Sugahara K
• 通讯作者: Sugahara K

Involvement of HNK-1 sulfotransferase in the biosynthesis of chondro itin sulfate and the GlcA(3-O-sulfate)-Gal-Gal-Xyl tetrasaccharide found in α-thrombomodulin from human urine

HNK-1 磺基转移酶参与硫酸软骨素和人尿 α-血栓调节蛋白中发现的 GlcA(3-O-硫酸盐)-Gal-Gal-Xyl 四糖的生物合成

• DOI: 10.1074/jbc.m111.279174
• 发表时间: 2011
• 期刊: J.Biol.Chem
• 作者: Maita H.;Tomita K. and Ariga H;相馬悠樹ほか;深水啓朗，鈴木晃 前野祐介，鈴木豊史，伴野和夫;Taishi Hashiguchi
• 通讯作者: Taishi Hashiguchi

ユニークな構造と機能をもつエビ頭部に由来するヘパラン硫酸

源自虾头的硫酸乙酰肝素具有独特的结构和功能

• 发表时间: 2012
• 作者: Shibata N;Toi S;Shibata T;Uchida K;Itabe H;Sawada T;Kawamata T;Okada Y;Uchiyama S;Kobayashi M.;羽磨紀城
• 通讯作者: 羽磨紀城

Involvement of the Hepatocyte Growth Factor Signaling Pathway in the Neuritogenic Activity of Chondroitin Sulfate Derived from Ray Fish Cartilage

肝细胞生长因子信号通路参与来自鳐鱼软骨的硫酸软骨素的神经炎活性

• 发表时间: 2010
• 作者: T. Hashiguchi;T. Kobayashi;D. Fongmoon;A. K. Shetty;S. Mizumoto;N. Miyamoto;T. Nakamura;S. Yamada;and K. Sugahara
• 通讯作者: and K. Sugahara