[人甲状腺肿甲状腺球蛋白的酶促去糖基化对其免疫化学性质的影响]

[Effects of enzymatic deglycosylation of human goiter thyroglobulin on its immunochemical properties].

作者信息

Grimaldi S, Fusco A, Olivieri A, Ioppolo A, Iacovacci P, Carlini F, Monaco F, Roche J

出版信息

C R Seances Soc Biol Fil. 1986;180(3):284-9.

Abstract

Thyroglobulin (Tg), isolated from soluble iodoproteins by ammonium sulphate fractionation, was enzymatically deglycosylated in vitro and analyzed by polyacrylamide gel electrophoresis, double immunodiffusion and non-commercial RIA. Carbohydrate and iodine content was chemically determined. By PAAGE deglycosylated Tg (dTg) showed the appearance of a major band in the 12S region and three slower migrating bands corresponding to higher aggregates than 19S Tg. In immunodiffusion by testing native and deglycosylated Tg against anti-native Tg antiserum it was shown the appearance of a spur of native on deglycosylated Tg. By RIA of native and deglycosylated Tg against anti-deglycosylated Tg antiserum it was shown a minor binding capacity of the anti-deglycosylated antibody against native Tg at high dilutions. The results demonstrate that the enzymatic deglycosylation release almost all the carbohydrates of goiter Tg and that the removal of the carbohydrates of Tg produces a loss of antigenic determinants of the molecule.

摘要

通过硫酸铵分级分离从可溶性碘蛋白中分离出甲状腺球蛋白（Tg），在体外对其进行酶促去糖基化，并通过聚丙烯酰胺凝胶电泳、双向免疫扩散和非商业性放射免疫分析（RIA）进行分析。化学测定碳水化合物和碘含量。通过聚丙烯酰胺凝胶电泳（PAAGE），去糖基化的Tg（dTg）在12S区域出现一条主要条带以及三条迁移速度较慢的条带，它们对应于比19S Tg更大的聚集体。在免疫扩散中，用天然Tg和去糖基化Tg对抗天然Tg抗血清进行检测，结果显示在去糖基化Tg上出现了天然Tg的刺突。通过用天然Tg和去糖基化Tg对抗去糖基化Tg抗血清进行RIA检测，结果显示在高稀释度下，抗去糖基化抗体对天然Tg的结合能力较弱。结果表明，酶促去糖基化几乎释放了甲状腺肿Tg的所有碳水化合物，并且去除Tg的碳水化合物会导致该分子抗原决定簇的丢失。