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Cell and molecular biology of enzymes involved in branch formation of mucin-type sugar chains

参与粘蛋白型糖链分支形成的酶的细胞和分子生物学

基本信息

批准号：
07458150
负责人：
TAKASAKI Seiichi
金额：
$ 4.16万
依托单位：
The University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1996
项目状态：
已结题

项目摘要

1. Regulation of oligosaccharide structures by core 2beta1,6 GlcNAc transferase (C2GnT) during lymphocyte activation : A human B lymphoblastic cell line, HuNS-1, was induced to express core 2 oligosaccharides by activation with IL-4. This induction was accompanied with increased expression of C2GnT activity and its mRNA.It is suggested, therefore, that the elevation of C2GnT may have some roles in the process of B cell activation.2. Expression of sialy Lewis X accompanied with branch formation of sugar chains and its corelation with adhesiveness to endothelial cells : The gene coding the enzyme involved in synthesis of polysialic acids was introduced to CHO cells expressing sialy Lewis X.This transfection resulted in the decreased expression of sialyl Lewis X and the reduced adhesion of the cells to E-selectin.3. Cloning of core 4beta1,6 GlcNAc transferase : The cDNA library was constructed from poly (A) + RNA of colon carcinoma LS174 cells. Then, several positive clones were obtained by cross hybridization, using homologous regions of the two beta1,6 GlcNAc transferases involved in synthesis of I antigenic determinant and core 2 oligosaccharide.4. Structure, function and biosynthesis of the sugar moieties expressed on the branched core : Cloning of CMP-NeuAc hydroxylase were successfully performed. The structures of sugar chains which may serve as ligands for T-cell adhesion molecule CD2 were characterized.

1.淋巴细胞活化过程中核心2 β 1，6 GlcNAc转移酶（C2 GnT）对寡糖结构的调节：通过IL-4活化诱导人B淋巴母细胞系HuNS-1表达核心2寡糖。结果：1.在诱导过程中伴随着C2 GnT活性及其mRNA表达的增加，提示C2 GnT的升高可能在B细胞活化过程中起一定作用. sialy刘易斯X的表达伴随糖链分支的形成及其与内皮细胞粘附的相关性：将编码多聚唾液酸合成酶的基因导入表达sialy刘易斯X的CHO细胞中，导致细胞对sialy刘易斯X的表达降低，并降低细胞对E-选择素的粘附。核心4 β 1，6 GlcNAc转移酶的克隆：从结肠癌LS 174细胞的poly（A）+ RNA构建cDNA文库。然后，利用参与I抗原决定簇和核心2寡糖合成的两种β 1，6 GlcNAc转移酶的同源区域进行交叉杂交，获得了多个阳性克隆.在分支核心上表达的糖部分的结构、功能和生物合成：成功地进行了CMP-NeuAc羟化酶的克隆。对可能作为T细胞粘附分子CD 2配体的糖链结构进行了表征。