Laurie S M, Cain C C, Lienhard G E, Castle J D
Department of Anatomy and Cell Biology, University of Virginia Health Sciences Center, Charlottesville 22908.
J Biol Chem. 1993 Sep 5;268(25):19110-7.
Secretory carrier membrane proteins (SCAMPs) mark the recycling system for the insulin-responsive glucose transporter, GluT4, in rat adipocytes. Anti-GluT4 and anti-SCAMP antibodies each immunoadsorbed vesicles containing both antigens from a low density microsomal fraction that is enriched in both antigens. The immunoadsorbed vesicles also contain VAMPs (synaptobrevins), synaptic vesicle membrane proteins. All three antigens were colocalized in low density microsomal vesicles from both basal and insulin-stimulated adipocytes. The SCAMPs have the same electrophoretic mobility as a major polypeptides detected in GluT4 vesicles. During insulin stimulation, 40% each of GluT4 and VAMPs redistribute from low density microsomes to the plasma membrane fraction; however, < 10% of the SCAMPs redistribute. Immunocytochemical staining of adipose tissue shows almost complete coincidence of SCAMPs and GluT4 in the basal state and extensive redistribution of both antigens to the cell periphery during insulin stimulation. Segregation of antigens during stimulation is not as distinct as observed by fractionation, although there are regions at the cell border where the SCAMPs appear more concentrated than GluT4. These data suggest that during insulin stimulation, in contrast to the behaviour of GluT4, SCAMPs remain tightly associated with the recycling system.
分泌载体膜蛋白(SCAMPs)标记了大鼠脂肪细胞中胰岛素反应性葡萄糖转运体GluT4的回收系统。抗GluT4抗体和抗SCAMP抗体各自免疫吸附了来自富含这两种抗原的低密度微粒体部分的同时含有这两种抗原的囊泡。免疫吸附的囊泡还含有囊泡相关膜蛋白(VAMPs,即突触小泡蛋白),突触小泡膜蛋白。在基础状态和胰岛素刺激的脂肪细胞的低密度微粒体囊泡中,所有这三种抗原都共定位。SCAMPs与在GluT4囊泡中检测到的一种主要多肽具有相同的电泳迁移率。在胰岛素刺激期间,GluT4和VAMPs各自有40%从低密度微粒体重新分布到质膜部分;然而,只有不到10%的SCAMPs重新分布。脂肪组织的免疫细胞化学染色显示,在基础状态下SCAMPs和GluT4几乎完全重合,在胰岛素刺激期间这两种抗原都广泛重新分布到细胞周边。尽管在细胞边界存在一些区域,其中SCAMPs似乎比GluT4更集中,但刺激期间抗原的分离并不像通过分级分离观察到的那样明显。这些数据表明,在胰岛素刺激期间,与GluT4的行为相反,SCAMPs与回收系统保持紧密关联。