Kijimoto-Ochiai S, Katagiri Y U, Hatae T, Okuyama H
Institute of Immunological Science, Hokkaido University, Sapporo, Japan.
Biochem J. 1989 Jan 1;257(1):43-9. doi: 10.1042/bj2570043.
The oligosaccharide chains of microheterogeneous bovine pancreatic DNAases were characterized by the lectin-nitrocellulose sheet method. The active fractions of the DNAases from column chromatography showed four major and several minor spots on a two-dimensional polyacrylamide gel. They were transferred on to nitrocellulose sheets and treated with glycosidases (neuraminidase, endo-beta-N-acetyl glucosaminidase H or F, or peptide N-glycosidase F) and treated with peroxidase-coupled lectins (concanavalin A, Ricinus communis agglutinin or wheat-germ agglutinin). From the results, the most probable oligosaccharide types were proposed to be as follows: the four major spots contained components which had high-mannose type or hybrid-type oligosaccharides, such as those susceptible to endo-beta-N-acetylglucosaminidase H. In addition, spot 1 contained a complex-type biantennary oligosaccharide without sialic acid and spot 3 contained a tri- or tetra-antennary complex-type oligosaccharide with sialic acid. The component corresponding to spot 2 had a hybrid-type oligosaccharide chain with a 'bisecting' acetylglucosamine, linked 1-4 to the beta-mannose residue of the trimannosyl core, and the component corresponding to spot 4 had a high-mannose-type oligosaccharide chain.
采用凝集素 - 硝酸纤维素膜法对微不均一性牛胰脱氧核糖核酸酶的寡糖链进行了表征。柱色谱法得到的脱氧核糖核酸酶活性组分在二维聚丙烯酰胺凝胶上显示出四个主要斑点和几个次要斑点。将它们转移到硝酸纤维素膜上，用糖苷酶（神经氨酸酶、内切β - N - 乙酰葡糖胺酶H或F或肽N - 糖苷酶F）处理，并用过氧化物酶偶联的凝集素（伴刀豆球蛋白A、蓖麻凝集素或麦胚凝集素）处理。根据结果，提出最可能的寡糖类型如下：四个主要斑点包含具有高甘露糖型或杂合型寡糖的组分，例如那些对内切β - N - 乙酰葡糖胺酶H敏感的组分。此外，斑点1包含一种不含唾液酸的复合型双天线寡糖，斑点3包含一种带有唾液酸的三天线或四天线复合型寡糖。与斑点2对应的组分具有一条带有“平分”型乙酰葡糖胺的杂合型寡糖链，该乙酰葡糖胺以1 - 4连接到三甘露糖核心的β - 甘露糖残基上，与斑点4对应的组分具有一条高甘露糖型寡糖链。
