Analysis of the biosynthetic mechanism and functions of glycosaminogycans by manipulating the genes encoding biosynthetic enzymes.

通过操纵编码生物合成酶的基因来分析糖胺聚糖的生物合成机制和功能。

• 批准号: 13470493
• 负责人: SUGAHARA Kazuyuki
• 金额: $ 7.81万
• 依托单位: Kobe Pharmaceutical University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-13470493/
• 关键词: glycosaminoglycan; heparan sulfate; chondroitin sulfate; glycosyltransferase; sulfotransferase; chondroitin synthase; chondroitin polymerizing factor; RNAi; ガン抑制遺伝子; EXT; プロテオグリカン; 線虫; 糖転位酵素; グルクロン酸転位酵素; ショウジョウバエ; シンデカン; グルクロン酸転移酵素; N-アセチ

(1) Human EXTL1 and EXTL3 were demonstrated for the first time to exhibit glycosyltransferase activities involved in heparan sulfate synthesis.(2) Human chondrotiin synthase was cloned for the first time.(3) It was demonstrated that acceptor core protein moieties in addition to an enzyme complex of human EXT1 and EXT2 for heparan sulfate polymerization.(4) Human chondroitin polymerizing factor required for chain polymerization of chondroitin was cloned for the first time.(5) Chondroitin/dermatan 4-O-sulfotransferases were cloned and their detailed substrate specificity was characterized.(6) It was found that one of the Gal residues in the GAG-protein linkage region tetrasaccharide is sulfated in the chondroitin sulfate chains but not in the heparan sulfate chains of a hybrid type proteoglycan syndecan-1.(7) The crystal structure of EXTL2, which exhibits GlcNAc transferase activity, was demonstrated for the first time.(8) Choncdoirin was demonstrated to be essential for the embryonic cell division of C.elegans.

(1)首次证明人EXTL 1和EXTL 3表现出参与硫酸乙酰肝素合成的糖基转移酶活性。(2)首次克隆了人软骨素合成酶。(3)证明了除了人EXT 1和EXT 2的酶复合物之外，受体核心蛋白部分用于硫酸乙酰肝素聚合。(4)首次克隆了软骨素链聚合所需的人软骨素聚合因子。(5)软骨素/皮肤素4-O-磺基转移酶进行了克隆，其详细的底物特异性进行了表征。(6)发现GAG-蛋白质连接区四糖中的Gal残基之一在硫酸软骨素链中被硫酸化，但在杂合型蛋白聚糖多配体蛋白聚糖-1的硫酸乙酰肝素链中未被硫酸化。(7)首次证明了具有GlcNAc转移酶活性的EXTL 2的晶体结构。(8)软骨素被证明是必不可少的胚胎细胞分裂的秀丽隐杆线虫。

项目成果

三上 雅之: "Specificities of three distinct human chondroitin/dermatan N-acetylgalactosamine 4-Ο-sulfotransferases demonstrated using partially desulfated dermatan sulfate as an acceptor : Implication of differential roles in dermatan sulfate biosynthesis."J.

Masayuki Mikami：“使用部分脱硫的硫酸皮肤素作为受体证明了三种不同的人软骨素/皮肤素 N-乙酰半乳糖胺 4-O-磺基转移酶的特异性：硫酸皮肤素生物合成中不同作用的含义。”

上野 百世: "Structural Characterization of Heparan Sulfate and Chondroitin Sulfate of Syndecan-1 Purified from Normal Murine Mammary Gland Epithelial Cells. Common Phosphorylation of Xylose and Differential Sulfation of Galactose in the Protein Linkage Region

Momoyo Ueno：“从正常鼠乳腺上皮细胞中纯化的 Syndecan-1 的硫酸乙酰肝素和硫酸软骨素的结构表征。蛋白质连接区域中木糖的常见磷酸化和半乳糖的差异硫酸化

Molecular cloning of a chondroitin polymerizing factor that cooperates with chondroitin synthase for chondroitin polymerization

• DOI: 10.1074/jbc.m302493200
• 发表时间: 2003-06-27
• 期刊: JOURNAL OF BIOLOGICAL CHEMISTRY
• 影响因子: 4.8
• 作者: Kitagawa, H;Izumikawa, T;Sugahara, K
• 通讯作者: Sugahara, K

Human glycosaminoglycan glucuronyltransferase I gene and a related processed pseudogene : genomic structure, chromosomal mapping, and characterization.

人糖胺聚糖葡萄糖醛酸转移酶 I 基因和相关的加工假基因：基因组结构、染色体作图和表征。

• 发表时间: 2001
• 期刊: Biochem.J. 358(3)
• 作者: Kitagawa;Hiroshi
• 通讯作者: Hiroshi

Specificities of three distinct human chondroitin/dermatan N-acetylgalactosamine 4-O-sulfotransferases demonstrated using partially desulfated dermatan sulfate as an acceptor -: Implication of differential roles in dermatan sulfate biosynthesis

• DOI: 10.1074/jbc.m306044200
• 发表时间: 2003-09-19
• 期刊: JOURNAL OF BIOLOGICAL CHEMISTRY
• 影响因子: 4.8
• 作者: Mikami, T;Mizumoto, S;Sugahara, K
• 通讯作者: Sugahara, K