Cushman S W, Goodyear L J, Pilch P F, Ralston E, Galbo H, Ploug T, Kristiansen S, Klip A
Experimental Diabetes, Metabolism, and Nutrition Section, NIDDK, National Institutes of Health, Bethesda, Maryland, USA.
Adv Exp Med Biol. 1998;441:63-71. doi: 10.1007/978-1-4899-1928-1_6.
Studies in mammalian cells have established the existence of numerous intracellular signaling cascades that are critical intermediates in the regulation of various biological functions. Over the past few years considerable research has shown that many of these signaling proteins are expressed in skeletal muscle. However, the detailed mechanisms involved in the regulation of glucose transporter (GLUT4) translocation from intracellular compartments to the cell surface membrane in response to insulin and contractions in skeletal muscle are not well understood. In the present essay we report three different approaches to unravel the GLUT4 translocation mechanism: 1. specific pertubation of the insulin and/or contraction signaling pathways; 2. characterization of the protein composition of GLUT4-containing vesicles with the expectation that knowledge of the constituent proteins of the vesicles may help in understanding their trafficking; 3. degree of co-immunolocalization of the GLUT4 glucose transporters with other membrane marker proteins assessed by immunofluorescense and electron microscopy.
在哺乳动物细胞中的研究已经证实存在众多细胞内信号级联反应,这些反应是调节各种生物学功能的关键中间体。在过去几年中,大量研究表明,许多此类信号蛋白在骨骼肌中表达。然而,骨骼肌中葡萄糖转运蛋白4(GLUT4)响应胰岛素和收缩从细胞内区室转运至细胞表面膜所涉及的详细机制尚不清楚。在本文中,我们报告了三种不同的方法来阐明GLUT4转运机制:1. 对胰岛素和/或收缩信号通路进行特异性干扰;2. 对含GLUT4囊泡的蛋白质组成进行表征,期望了解囊泡的组成蛋白可能有助于理解其运输过程;3. 通过免疫荧光和电子显微镜评估GLUT4葡萄糖转运蛋白与其他膜标记蛋白的共免疫定位程度。