Christie D L, Palmer D J
Department of Biochemistry, University of Auckland, New Zealand.
Biochem J. 1990 Aug 15;270(1):57-61. doi: 10.1042/bj2700057.
Chromaffin-granule membranes were separated into insoluble and soluble fractions after extraction with lithium di-iodosalicylate (LDIS). These fractions were characterized by one- and two-dimensional gel electrophoresis, and glycoproteins were detected after electroblotting with peroxidase-labelled concanavalin A and wheat-germ agglutinin (WGA). The LDIS-insoluble fraction contained components identified as glycoproteins III, H, J and K (carboxypeptidase H). Microsequence analysis indicated that component J is an N-terminally extended form of glycoprotein K. A major glycoprotein, GpII (Mr 80,000-100,000), present in the LDIS-soluble fraction was purified by affinity chromatography on WGA-Sepharose. This was characterized by one- and two-dimensional gel electrophoresis with Coomassie Blue staining, by amino acid analysis and automated N-terminal sequence analysis. Extraction of chromaffin-granule membranes with LDIS is a simple and rapid procedure that facilitates studies concerned with the structure and function of membrane glycoproteins from these and other secretory granules.
用二碘水杨酸锂（LDIS）提取后，嗜铬粒细胞膜被分离成不溶性和可溶性部分。这些部分通过一维和二维凝胶电泳进行表征，在用过氧化物酶标记的伴刀豆球蛋白A和麦胚凝集素（WGA）进行电印迹后检测糖蛋白。LDIS不溶性部分包含被鉴定为糖蛋白III、H、J和K（羧肽酶H）的成分。微序列分析表明成分J是糖蛋白K的N端延伸形式。通过在WGA-琼脂糖上进行亲和层析，纯化了存在于LDIS可溶性部分中的一种主要糖蛋白GpII（分子量80,000 - 100,000）。通过考马斯亮蓝染色的一维和二维凝胶电泳、氨基酸分析和自动N端序列分析对其进行表征。用LDIS提取嗜铬粒细胞膜是一种简单快速的方法，有助于研究这些及其他分泌颗粒膜糖蛋白的结构和功能。
