权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Study on the biological function of glycoconjugates

复合糖的生物学功能研究

基本信息

批准号：
12680610
负责人：
HABUCHI Osami
金额：
$ 2.3万
依托单位：
Aichi University of Education
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2001
项目状态：
已结题

项目摘要

(1) We cloned mouse and human chondroitin 4-sulfotransferase gene, characterized the protein expressed from the gene and determined expression pattern in various tissues. We also mapped the gene in human chromosome.(2) We cloned and characterized human GalNAc 4-sulfotransferase that strongly expressed in human pituitary grand. We also cloned mouse GalNAc 4-sulfotransferase I and II and determined in various tissues by Northern blot and RT-PCR.(3) We expressed NSIST, which was cloned from Torpedo electric organ, in COS-7 cells and found that substrate specificity of NSIST was very similar to that of chondroitin 6-sulfotransferase.(4) Af-Acetylgalactosamine 4-sulfate 6-O-sulfotransferase (GalNAc4S-6ST)was purified to apparent homogeneity from the squid cartilage. The purified enzyme transferred sulfate from PAPS to chondroitin sulfate A, chondroitin sulfate C and dermatan sulfate. The transfer of sulfate to chondroitin sulfate A and dermatan sulfate occurred mainly at position 6 of the i … More nternal Af-acetylgalactosamine 4-sulfate residues. When a trisaccharide or a pentasaccharide having sulfate groups at position 4 of W-acetylgalactosamine residues were used as acceptors, efficient sulfation of position 6 at nonreducing terminal N-acetylgalactosamine 4-sulfate residue was observed. We cloned and characterized human GalNAc4S-6ST. The strategy for identification of human GalNAc4S-6ST consisted of: 1) determination of the amino acid sequences of peptides derived from the purified squid GalNAc4S-6ST, 2) amplification of squid DNA by PCR, and 3) homology search using the amino acid sequence deduced from the squid DNA. The recombinant protein expressed from the human GalNAc4S-6ST cDNA transferred sulfate from PAPS to position 6 of the nonreducing terminal and internal GalNAc (4SO_4) residues contained in chondroitin sulfate A and dermatan sulfate. When a trisaccharide and a pentasaccharide having sulfate groups at position 4 of N-acetylgalactosamine residues were used as acceptors, only nonreducing terminal GalNAc (4SO_4) residues were sulfated. The nucleotide sequence of the human GalNAc4S-6ST cDNA was nearly identical to the sequence of human B cell RAG-associated gene. Less

(1)我们克隆了小鼠和人软骨素4-磺基转移酶基因，表征了从该基因表达的蛋白质，并确定了在各种组织中的表达模式。我们还定位了该基因在人类染色体上的位置。(2)我们克隆了人GalNAc 4-磺基转移酶，并对其进行了鉴定。我们还克隆了小鼠GalNAc 4-磺基转移酶I和II，并通过北方印迹和RT-PCR在各种组织中进行了测定。(3)我们在COS-7细胞中表达了从电鳐电器官中克隆的NSIST，发现NSIST的底物特异性与软骨素6-磺基转移酶的底物特异性非常相似。(4)从鱿鱼软骨中纯化了α f-乙酰半乳糖胺4-硫酸6-O-磺基转移酶（GalNAc 4S-6ST）。纯化的酶将硫酸盐从PAPS转移到硫酸软骨素A、硫酸软骨素C和硫酸皮肤素。硫酸盐向硫酸软骨素A和硫酸皮肤素的转移主要发生在I型胶原的6位。 ...更多信息内部N-乙酰半乳糖胺4-硫酸盐残基。当使用在N-乙酰半乳糖胺残基的4位上具有硫酸酯基团的三糖或五糖作为受体时，观察到在非还原性末端N-乙酰半乳糖胺4-硫酸酯残基的6位上的有效硫酸化。我们克隆并鉴定了人GalNAc 4S-6ST。鉴定人GalNAc 4S-6ST的策略包括：1）确定来自纯化的鱿鱼GalNAc 4S-6ST的肽的氨基酸序列，2）通过PCR扩增鱿鱼DNA，和3）使用从鱿鱼DNA推导的氨基酸序列进行同源性搜索。用人GalNAc 4S-6ST cDNA表达的重组蛋白将PAPS中的硫酸根转移到硫酸软骨素A和硫酸皮肤素的非还原性末端和内部GalNAc（4SO_4）残基的6位。当N-乙酰半乳糖胺残基的4位具有硫酸基的三糖和五糖用作受体时，仅非还原性末端GalNAc（4SO_4）残基被硫酸化。人GalNAc 4S-6ST cDNA的核苷酸序列与人B细胞RAG相关基因的序列几乎相同。少