CHARACTERIZATION AND TOPOGRAPHY OF THE GLYCOPROTEINS OF ADRENAL CHROMAFFIN GRANULES
CHARACTERIZATION AND TOPOGRAPHY OF THE GLYCOPROTEINS OF ADRENAL CHROMAFFIN GRANULES
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DOI：
10.1111/j.1471-4159.1979.tb04507.x
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发表时间：
1979-01-01
影响因子：
4.7
通讯作者：
WINKLER, H
中科院分区：
文献类型：
--
作者：
HUBER, E;KONIG, P;WINKLER, H
采用2个聚丙烯酰胺凝胶电泳系统对牛嗜铬颗粒膜的糖蛋白进行了表征。五个组分 (I-V) 的未还原形式的表观分子量范围为 45,000-150,000。糖蛋白I被鉴定为多巴胺β-羟化酶。在这些糖蛋白中（除了组分 IV），有 4 种显然也存在于猪和马嗜铬颗粒的膜中。嗜铬颗粒的可溶性蛋白质至少含有3种糖蛋白。仅糖蛋白I（多巴胺β-羟化酶）存在于可溶物和嗜铬颗粒的膜中。使用凝集素的亲和层析表明，在可溶性蛋白质中，只有多巴胺β-羟化酶被伴刀豆球蛋白[Con]A吸附，而这些蛋白质均不与麦芽凝集素和蓖麻凝集素反应。包括多巴胺β-羟化酶和糖蛋白II在内的三种膜蛋白作为主要成分被Con A吸附，而麦芽凝集素仅结合成分II和少量成分III。通过 EM，证明刀豆蛋白 A 不与完整的嗜铬颗粒结合，而钌红和阳离子化铁蛋白却可以。半乳糖氧化酶处理后的同位素标记表明，至少主要糖蛋白的碳水化合物部分存在于面向内容物的颗粒膜的内侧。
The glycoproteins of the membranes of bovine chromaffin granules were characterized by 2 polyacrylamide gel electrophoresis systems. Five components (I-V) were demonstrated with apparent MW ranging in the unreduced form from 45,000-150,000. Glycoprotein I was identified as the enzyme dopamine .beta.-hydroxylase. Of these glycoproteins (with the exception of component IV), 4 were apparently also present in the membranes of pig and horse chromaffin granules. The soluble protein of chromaffin granules contained at least 3 glycoproteins. Only glycoprotein I (dopamine .beta.-hydroxylase) was present in the soluble content and in the membranes of chromaffin granules. Affinity chromatography with lectins demonstrated that from the soluble proteins only dopamine .beta.-hydroxylase was adsorbed by concanavalin [Con] A, whereas none of these proteins reacted with wheat germ lectin and Ricinus communis agglutinin. Three membrane proteins including dopamine .beta.-hydroxylase and glycoprotein II as major components were adsorbed by Con A, whereas wheat germ lectin bound only component II and a small amount of component III. By EM it was demonstrated that Con A did not bind to intact chromaffin granules, whereas ruthenium red and cationized ferritin did. Isotope labeling after galactose oxidase treatment revealed that at least the carbohydrate portion of the major glycoproteins was present on the inner side of the granule membranes facing the content.