Mastick C C, Aebersold R, Lienhard G E
Department of Biochemistry, Dartmouth Medical School, Hanover, New Hampshire 03755.
J Biol Chem. 1994 Feb 25;269(8):6089-92.
GLUT4-enriched vesicles from fat and muscle cells contain a major protein of 165 kDa. In the present study we have prepared GLUT4 vesicles from a large number of rat adipocytes, isolated the 165-kDa protein by SDS-gel electrophoresis, and obtained the sequences of four tryptic peptides. Comparison of the sequences with those in protein data banks showed that the 165-kDa protein had not previously been sequenced. An antibody preparation against one of the peptides immunoblotted the 165-kDa protein. By this means it was found that the 165-kDa protein was concentrated in GLUT4 vesicles, and that insulin treatment of rat adipocytes caused a translocation of the protein to the plasma membrane that paralleled the translocation of GLUT4.
来自脂肪和肌肉细胞的富含葡萄糖转运蛋白4（GLUT4）的囊泡含有一种主要的165 kDa蛋白质。在本研究中，我们从大量大鼠脂肪细胞中制备了GLUT4囊泡，通过SDS凝胶电泳分离出165 kDa蛋白质，并获得了四个胰蛋白酶肽段的序列。将这些序列与蛋白质数据库中的序列进行比较，结果表明此前尚未对该165 kDa蛋白质进行测序。针对其中一个肽段制备的抗体对165 kDa蛋白质进行了免疫印迹分析。通过这种方法发现，165 kDa蛋白质集中在GLUT4囊泡中，并且对大鼠脂肪细胞进行胰岛素处理会导致该蛋白质向质膜的转运，这与GLUT4的转运情况相似。
