肠道微粒体中岩藻糖基转移酶系统的研究。

Study of the fucosyltransferase system in intestinal microsomes.

作者信息

Martin A, Biol M C, Broquet P, Richard M, Louisot P

机构信息

Department of General and Medical Biochemistry, INSERM-CNRS U 189, Lyon-Sud Medical School, Oullins, France.

出版信息

Enzyme. 1988;39(1):17-27. doi: 10.1159/000469090.

DOI:10.1159/000469090

PMID:3345733

Abstract

Fucosyltransferase activity of rat small intestine microsomes is solubilized by 0.5% Triton X-100. The solubilized activity can be purified up to 8,300-fold using DEAE-cellulose and affinity chromatography on GDP-Sepharose. At this step, chromatography on Sephadex G15 separates different specificities: N-acetylglucosaminide-alpha-(1,3)-fucosyltransferase acting on asialoserotransferrin, and galactoside-alpha-(1,2)-fucosyltransferase acting on O-glycans of asialofetuin. The use of small saccharidic acceptors also indicates the presence of a N-acetylglucosaminide-alpha-(1,4)-fucosyltransferase and of a very weak glucose-alpha-(1,3)-fucosyltransferase activity. These activities are tightly bound to concanavalin A-Sepharose, suggesting that they are supported by N-glycosylproteins.

摘要

大鼠小肠微粒体的岩藻糖基转移酶活性可被0.5% Triton X - 100溶解。使用DEAE - 纤维素和GDP - Sepharose亲和层析可将溶解后的活性纯化至8300倍。在此步骤中，用Sephadex G15层析可分离出不同的特异性：作用于去唾液酸血清转铁蛋白的N - 乙酰葡糖胺 - α - (1,3) - 岩藻糖基转移酶，以及作用于去唾液酸胎球蛋白O - 聚糖的半乳糖苷 - α - (1,2) - 岩藻糖基转移酶。使用小分子糖类受体也表明存在N - 乙酰葡糖胺 - α - (1,4) - 岩藻糖基转移酶和非常微弱的葡萄糖 - α - (1,3) - 岩藻糖基转移酶活性。这些活性与伴刀豆球蛋白A - Sepharose紧密结合，表明它们由N - 糖基化蛋白支持。