牛角膜硫酸角质素蛋白聚糖中多糖链与核心蛋白之间连接区域的特性研究。

Studies on the characterization of the linkage-region between polysaccharide chain and core protein in bovine corneal proteokeratan sulfate.

Keller R, Stein T, Stuhlsatz H W, Greiling H, Ohst E, Müller E, Scharf H D

Hoppe Seylers Z Physiol Chem. 1981 Mar;362(3):327-36. doi: 10.1515/bchm2.1981.362.1.327.

Abstract

A new method of enrichment of the linkage-region in corneal proteokeratan sulfate is described, which consists of desulfation of peptidokeratan sulfate, followed by chromatography on Con A-Sepharose 4B and enzymatic degradation with beta-D galactosidase and beta-N-acetyl-D-glucosaminidase. 2) After permethylation, hydrolysis, reduction with sodium borohydrid and acetylation gas chromatography/mass spectrometry analyses were performed. The followings products could be detected as their peracetates: 2,3,4-tri-O-methylfucitol; 2,3,4,6-tetra-O-methylmannitol; 3,4,6-tri-O-methylmannitol; 2,4-di-O-methylmannitol; 2,3,4,6-tetra-O-methylgalactitol; 2,4,6-tri-O-methylgalactitol; 2,4-di-O-methylgalactitol. 3) The results point to the presence of a branched linkage region in the proteokeratan sulfate molecule with one mannose as the branching point and two mannose residues as the starting point of two disaccharide chains.

摘要

描述了一种富集角膜蛋白硫酸角质素中连接区的新方法，该方法包括对肽硫酸角质素进行脱硫，然后在伴刀豆球蛋白A - 琼脂糖4B上进行色谱分析，并用β - D - 半乳糖苷酶和β - N - 乙酰 - D - 葡糖胺酶进行酶解。2) 经全甲基化、水解、硼氢化钠还原和乙酰化后，进行气相色谱/质谱分析。以下产物可作为其全乙酸酯被检测到：2,3,4 - 三 - O - 甲基岩藻糖醇；2,3,4,6 - 四 - O - 甲基甘露糖醇；3,4,6 - 三 - O - 甲基甘露糖醇；2,4 - 二 - O - 甲基甘露糖醇；2,3,4,6 - 四 - O - 甲基半乳糖醇；2,4,6 - 三 - O - 甲基半乳糖醇；2,4 - 二 - O - 甲基半乳糖醇。3) 结果表明，蛋白硫酸角质素分子中存在一个分支连接区，其中一个甘露糖作为分支点，两个甘露糖残基作为两条二糖链的起始点。