固定化乳过氧化物酶介导的淋巴细胞表面蛋白和糖蛋白的放射性碘化

Radioiodination of surface proteins and glycoproteins of lymphocytes by immobilized lactoperoxidase.

作者信息

Koch N, Haustein D

出版信息

J Immunol Methods. 1981;41(2):163-71. doi: 10.1016/0022-1759(81)90240-4.

Abstract

Radioiodination of lymphocyte surface proteins employing soluble lactoperoxidase was found to be unsatisfactory for the quantitative isolation and characterization of cell surface glycoproteins: either the glycoproteins were contaminated by self-iodinated lactoperoxidase or were in part removed by the washing step following the radioiodination procedure. Therefore, optimal conditions for cell surface radioiodination by lactoperoxidase covalently linked to Sepharose 4B (lacto-beads) were worked out. By the employment of immobilized lactoperoxidase, the enzyme could easily be removed following solubilization of the radiolabelled cells by a simple washing step. Radioiodinated cell surface proteins and glycoproteins were obtained without any loss as shown by SDS polyacrylamide gel electrophoresis.

摘要

利用可溶性乳过氧化物酶对淋巴细胞表面蛋白进行放射性碘化，在定量分离和鉴定细胞表面糖蛋白方面并不理想：要么糖蛋白被自身碘化的乳过氧化物酶污染，要么在放射性碘化步骤后的洗涤过程中部分被去除。因此，研究出了用与琼脂糖4B共价连接的乳过氧化物酶（乳珠）进行细胞表面放射性碘化的最佳条件。通过使用固定化乳过氧化物酶，在通过简单的洗涤步骤使放射性标记细胞溶解后，可以很容易地将酶去除。如SDS聚丙烯酰胺凝胶电泳所示，获得了无任何损失的放射性碘化细胞表面蛋白和糖蛋白。