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The t-SNAREs syntaxin4 and SNAP23 but not v-SNARE VAMP2 are indispensable to tether GLUT4 vesicles at the plasma membrane in adipocyte
Authors:Takayuki Kawaguchi  Hajime Kanda  Sanshiro Tateya  Masato Kasuga
Institution:a Division of Diabetes, Metabolism, and Endocrinology, Department of Internal Medicine, Kobe University Graduate School of Medicine, Japan
b Research Institute, International Medical Center of Japan, Japan
Abstract:SNARE proteins (VAMP2, syntaxin4, and SNAP23) have been thought to play a key role in GLUT4 trafficking by mediating the tethering, docking and subsequent fusion of GLUT4-containing vesicles with the plasma membrane. The precise functions of these proteins have remained elusive, however. We have now shown that depletion of the vesicle SNARE (v-SNARE) VAMP2 by RNA interference in 3T3-L1 adipocytes inhibited the fusion of GLUT4 vesicles with the plasma membrane but did not affect tethering of the vesicles to the membrane. In contrast, depletion of the target SNAREs (t-SNAREs) syntaxin4 or SNAP23 resulted in impairment of GLUT4 vesicle tethering to the plasma membrane. Our results indicate that the t-SNAREs syntaxin4 and SNAP23 are indispensable for the tethering of GLUT4 vesicles to the plasma membrane, whereas the v-SNARE VAMP2 is not required for this step but is essential for the subsequent fusion event.
Keywords:GLUT4  glucose transporter 4  t- and v-SNARE  target and vesicle soluble N-ethylmaleimide-sensitive factor-attachment protein (SNAP) receptor  respectively  VAMP2  vesicle-associated membrane protein 2  GFP  green fluorescent protein  DMEM  Dulbecco&rsquo  s modified Eagle&rsquo  s medium  PBS  phosphate-buffered saline  PE  phycoerythrin  RNAi  RNA interference  siRNA  small interfering RNA
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