通过凝集素亲和柱色谱法以及与抗胰高血糖素抗体反应对化学氚标记的大分子胰高血糖素免疫反应性蛋白种类进行表征。

Characterization of a chemically tritiated large molecular weight glucagon immunoreactive protein species by lectin-affinity column chromatography and reaction with anti-glucagon antibodies.

作者信息

Tung A K, Siu K P

出版信息

Biochem Biophys Res Commun. 1984 Dec 14;125(2):524-31. doi: 10.1016/0006-291x(84)90571-0.

Abstract

High molecular weight glucagon immunoreactive material, obtained by gel-filtration (in the presence of 6 M guanidine hydrochloride) of fetal bovine pancreatic extracts, was tritiated by reductive methylation. Concanavalin-A-Sepharose column chromatography of the radiolabeled preparation yielded a discrete Concanavalin-A-reactive, alpha-methyl-mannoside-displaceable radioactive peak, coinciding with the glucagon immunoreactive peak. Submission of the Con-A-reactive material to wheat germ agglutinin-Sepharose column chromatography yielded a lectin-reactive, N-acetyl-glucosamine-displaceable radioactive peak, coinciding with the glucagon immunoreactive peak. The tritiated Con-A-reactive component interacted specifically with anti-glucagon antibodies. Sephacryl S-200 gel-filtration (in the presence of guanidine hydrochloride) dissociated a approximately 40 kDa radioactive species from the antibody-antigen complex. These data provide direct evidence for the existence of a large molecular weight glycosylated glucagon-related protein species from the fetal bovine pancreas.

摘要

通过对胎牛胰腺提取物进行凝胶过滤（在6M盐酸胍存在下）获得的高分子量胰高血糖素免疫反应性物质，经还原甲基化进行氚标记。对放射性标记制剂进行伴刀豆球蛋白A-琼脂糖柱色谱分析，得到一个离散的伴刀豆球蛋白A反应性、α-甲基甘露糖苷可置换的放射性峰，与胰高血糖素免疫反应性峰一致。将伴刀豆球蛋白A反应性物质进行麦胚凝集素-琼脂糖柱色谱分析，得到一个凝集素反应性、N-乙酰葡糖胺可置换的放射性峰，与胰高血糖素免疫反应性峰一致。氚标记的伴刀豆球蛋白A反应性成分与抗胰高血糖素抗体特异性相互作用。Sephacryl S-200凝胶过滤（在盐酸胍存在下）从抗体-抗原复合物中解离出一种约40kDa的放射性物质。这些数据为胎牛胰腺中存在一种高分子量糖基化胰高血糖素相关蛋白物质提供了直接证据。