GP160, A TISSUE-SPECIFIC MARKER FOR INSULIN-ACTIVATED GLUCOSE-TRANSPORT
GP160, A TISSUE-SPECIFIC MARKER FOR INSULIN-ACTIVATED GLUCOSE-TRANSPORT
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DOI：
10.1073/pnas.91.17.8017
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发表时间：
1994-08-16
影响因子：
11.1
通讯作者：
PILCH, PF
中科院分区：
文献类型：
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作者：
KANDROR, KV;PILCH, PF
我们已经分离并部分测序了一种M(r), 160,000的糖蛋白，其进出脂肪细胞表面的循环速率被胰岛素提高，其方式显然与胰岛素对GLUT4循环的影响相同。根据该蛋白的序列，我们制备了针对该蛋白的抗肽抗体gp160。该抗体识别M(r) 160,000蛋白，其亚细胞分布与GLUT4相同。这是通过三个独立的标准来确定的：(i)从暴露于胰岛素或未暴露于胰岛素的细胞中分离的脂肪细胞膜进行Western blotting， （ii）用抗glut4抗体吸附囊泡，然后进行Western blotting，以及（iii）通过蔗糖速度和密度梯度分离微粒体囊泡。通过这三个标准，GLUT4和gp160在大鼠脂肪细胞中完全共定位。此外，gp160仅能在脂肪、心肌和骨骼肌中检测到，其他组织均未检测到。因此，gp160是生理上重要的、胰岛素敏感的葡萄糖转运的额外标记物，其在蛋白质和DNA水平上的进一步研究可能揭示胰岛素激活的GLUT4易位的机制细节以及GLUT4和gp160的组织特异性表达的信息。
We have isolated and partially sequenced a M(r), 160,000 glycoprotein whose rate of cycling to and from the adipocyte cell surface is enhanced by insulin in a manner apparently identical to the effect of insulin on GLUT4 cycling. Based on the protein sequence, we have prepared an antipeptide antibody against this protein, gp160. The antibody recognizes a M(r) 160,000 protein whose subcellular distribution is identical to that of GLUT4. This was determined by three separate criteria: (i) Western blotting of fractionated adipocyte membranes from cells exposed to insulin or not, (ii) adsorption of vesicles with anti-GLUT4 antibodies followed by Western blotting, and (iii) separation of microsomal vesicles by sucrose velocity and density gradients. By all three criteria, GLUT4 and gp160 are completely colocalized in rat fat cells. Moreover, gp160 can be detected by Western blot only in fat and cardiac and skeletal muscles and was absent from all other tissues tested. Thus, gp160 is an additional marker for physiologically important, insuln-sensitive glucose transport, Its further study at the protein and DNA level may reveal information about the mechanistic details of insulin-activated GLUT4 translocation as well as information concerning the tissue-specific expression of GLUT4 and gp160.