Association of N-ethylmaleimide sensitive fusion (NSF) protein and soluble NSF attachment proteins-alpha and -gamma with glucose transporter-4-containing vesicles in primary rat adipocytes

Endocrinology
C C Mastick, A L Falick

Abstract

To investigate the role of N-ethylmaleimide sensitive fusion protein (NSF) and soluble NSF attachment proteins (SNAP)-containing fusion complexes in glucose transporter-4 (GLUT4) membrane trafficking, the subcellular distributions of NSF, alpha-SNAP, and gamma-SNAP in primary rat adipocytes were determined. A large fraction of the NSF and SNAPs were associated with intracellular membranes, distributed between the low-density microsomes (LDM) and high-density microsomes. Very little of the NSF and SNAPs were associated with the plasma membrane fraction. This distribution did not change after insulin stimulation. Approximately 75% of the NSF and SNAPs in the LDM fraction were coimmunoprecipitated with 85% of the GLUT4 and 60% of the vesicle associated membrane proteins (VAMPs; synaptobrevins) VAMP-2 and cellubrevin in anti-GLUT4 immunoadsorptions. In contrast to NSF and the SNAPs, the beta-coatomer protein (beta-COP) found in the LDM fraction was excluded from GLUT4 vesicles. When LDM fractions were solubilized with Thesit (octaethylene glycol dodecyl ether) or Triton X-100, approximately 40% of the alpha-SNAP was colocalized with NSF on glycerol gradients in large (approximately 20S), ATP-sensitive complexes. VAMP-2 and cellubre...Continue Reading

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Citations

Mar 3, 1999·The Biochemical Journal·J F St-DenisP A Roche
Dec 28, 1999·Proceedings of the National Academy of Sciences of the United States of America·G InoueC R Kahn
Sep 23, 2009·The Journal of Endocrinology·Yi Lin, Zhongjie Sun
Nov 25, 2005·Molecular Medicine·Louise ChangAlan R Saltiel
May 13, 2003·Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences·Fabrice DupratJohn Isaac
Jul 5, 2003·Physiological Reviews·Pamela L Tuma, Ann L Hubbard
Jun 24, 2008·The Biochemical Journal·Hilal ZaidAmira Klip
Sep 13, 2005·The Journal of Biological Chemistry·Mark LaranceDavid E James
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