前循环酸性重复蛋白的简易纯化及唾液酸化糖基磷脂酰肌醇膜锚定的证明
A simple purification of procyclic acidic repetitive protein and demonstration of a sialylated glycosyl-phosphatidylinositol membrane anchor.
作者信息
Ferguson M A, Murray P, Rutherford H, McConville M J
机构信息
Department of Biochemistry, University of Dundee, Scotland, U.K.
出版信息
Biochem J. 1993 Apr 1;291 ( Pt 1)(Pt 1):51-5. doi: 10.1042/bj2910051.
Abstract
The procyclic acidic repetitive protein is the major cell-surface glycoprotein of the insect-dwelling procyclic forms of the Trypanosoma brucei species of African trypanosomes. The glycoprotein contains an acidic Glu-Pro repeat domain, a glycosyl-phosphatidylinositol membrane anchor and a putative asparagine glycosylation site. In this paper we describe a rapid purification scheme for this glycoprotein, using solvent extraction and hydrophobic interaction chromatography, and a partial characterization of the glycosylphosphatidylinositol membrane anchor. The carbohydrate composition of the anchor is extremely unusual; it contains on average nine GlcNAc, nine Gal, and five sialic acid residues. This is the first description of such a heavily substituted and negatively charged anchor. A comparison between the trypanosome procyclic surface and the Leishmania promastigote surface is also presented.
摘要
前循环酸性重复蛋白是非洲锥虫布氏锥虫昆虫寄生前循环形式的主要细胞表面糖蛋白。该糖蛋白包含一个酸性的Glu-Pro重复结构域、一个糖基磷脂酰肌醇膜锚和一个假定的天冬酰胺糖基化位点。在本文中，我们描述了一种使用溶剂萃取和疏水相互作用色谱法对该糖蛋白进行快速纯化的方案，以及对糖基磷脂酰肌醇膜锚的部分特性分析。该锚的碳水化合物组成极为罕见；平均含有九个N-乙酰葡糖胺、九个半乳糖和五个唾液酸残基。这是对如此高度取代且带负电荷的锚的首次描述。还对锥虫前循环表面与利什曼原虫前鞭毛体表面进行了比较。
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Cell-surface anchoring of proteins via glycosyl-phosphatidylinositol structures.通过糖基磷脂酰肌醇结构实现蛋白质的细胞表面锚定。
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