人胎盘碱性磷酸酶糖基磷脂酰肌醇膜锚的结构
Structure of the glycosylphosphatidylinositol membrane anchor of human placental alkaline phosphatase.
作者信息
Redman C A, Thomas-Oates J E, Ogata S, Ikehara Y, Ferguson M A
机构信息
Department of Biochemistry, University of Dundee, U.K.
出版信息
Biochem J. 1994 Sep 15;302 ( Pt 3)(Pt 3):861-5. doi: 10.1042/bj3020861.
Abstract
The glycosylphosphatidylinositol membrane anchor of human placental alkaline phosphatase was isolated by exhaustive proteolysis followed by hydrophobic interaction chromatography. The resulting glycosylphosphatidylinositol-peptide was subjected to compositional analysis and chemical and enzymic modifications. The neutral-glycan fraction, prepared by dephosphorylation followed by HNO2 deamination and reduction, was sequenced using exoglycosidases and acetolysis. The phosphatidylinositol moiety was analysed by fast-atom bombardment mass spectrometry and gas chromatography-mass spectrometry. Taken together the data suggest the structure, Thr-Asp-ethanolamine-PO4-Man alpha 1-2Man alpha 1-6Man alpha 1-4GlcN-(sn-1-O- alkyl-2-O-acylglycerol-3-PO4-1-myo-D-inositol), which contains an additional ethanolamine phosphate group at an unknown position.
摘要
通过彻底的蛋白酶解，随后进行疏水相互作用色谱法，分离出了人胎盘碱性磷酸酶的糖基磷脂酰肌醇膜锚。对所得的糖基磷脂酰肌醇 - 肽进行了组成分析以及化学和酶促修饰。通过去磷酸化，然后进行亚硝酸脱氨和还原制备的中性聚糖部分，使用外切糖苷酶和乙酰解进行测序。通过快原子轰击质谱法和气相色谱 - 质谱法分析磷脂酰肌醇部分。综合这些数据表明其结构为苏氨酸 - 天冬氨酸 - 乙醇胺 - 磷酸 - 甘露糖α1 - 2甘露糖α1 - 6甘露糖α1 - 4葡糖胺 -（sn - 1 - O - 烷基 - 2 - O - 酰基甘油 - 3 - 磷酸 - 1 - 肌醇），在一个未知位置含有一个额外的乙醇胺磷酸基团。
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