Vesicle-associated membrane protein 2 plays a specific role in the insulin-dependent trafficking of the facilitative glucose transporter GLUT4 in 3T3-L1 adipocytes

被引:125
作者
Martin, LB
Shewan, A
Millar, CA
Gould, GW
James, DE [1 ]
机构
[1] Univ Queensland, Ctr Cellular & Mol Biol, St Lucia, Qld 4072, Australia
[2] Univ Queensland, Dept Physiol & Pharmacol, St Lucia, Qld 4072, Australia
[3] Univ Glasgow, Div Biochem & Mol Biol, Glasgow G12 8QQ, Lanark, Scotland
基金
英国惠康基金;
关键词
D O I
10.1074/jbc.273.3.1444
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Vesicle-associated membrane protein 2 (VAMP2) has been implicated in the insulin-regulated trafficking of GLUT4 in adipocytes. It has been proposed that VAMP2 co-localizes with GLUT4 in a postendocytic storage compartment (Martin, S., Tellam, J., Livingstone, C., Slot, J. W., Gould, G. W., and James, D. E. (1996) J. Cell Biol. 134, 625-635), suggesting that it may play a role distinct from endosomal v-SNAREs (soluble N-ethylmaleimide-sensitive factor attachment protein receptors) such as cellubrevin that are also expressed in adipocytes, The present study examines the effects of recombinant glutathione S-transferase (GST) fusion proteins encompassing the entire cytoplasmic tails of VAMP1, VAMP2, and cellubrevin on insulin-stimulated GLUT4 translocation in streptolysin O permeabilized 3T3-L1 adipocytes, GST-VAMPS inhibited insulin-stimulated GLUT4 trans location by similar to 35%, whereas GST-VAMP1 and GST-cellubrevin were without effect, A synthetic peptide corresponding to the unique N terminus of VAMP2 also inhibited insulin-stimulated GLUT4 translocation in a dose-dependent manner, This peptide had no effect on either guanosine 5'-3-O-(thio)triphosphate-stimulated GLUT4 translocation or on insulin-stimulated GLUT1 translocation, These results imply that GLUT4 and GLUT1 may undergo insulin stimulated translocation to the cell surface from separate intracellular compartments, To confirm this, adipocytes were incubated with a transferrin-horseradish peroxidase conjugate to fill the itinerant endocytic system after which cells were incubated with H2O2 and diaminobenzidine. This treat ment completely blocked insulin-stimulated movement of GLUT1, whereas in the case of GLUT4, movement to the surface was delayed but still reached similar levels to that observed in insulin-stimulated control cells after 30 min, These results suggest that the N terminus of VAMP2 plays a unique role in the insulin-dependent recruitment of GLUT4 from its intracellular storage compartment to the cell surface.
引用
收藏
页码:1444 / 1452
页数:9
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