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Am J Physiol Endocrinol Metab 287: E547-E552, 2004. First published May 27, 2004; doi:10.1152/ajpendo.00571.2003
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V-type ATPase is involved in biogenesis of GLUT4 vesicles

Marina Malikova,* Jun Shi,* and Konstantin V. Kandror

Boston University School of Medicine, Boston, Massachusetts 02118

Submitted 16 December 2003 ; accepted in final form 11 May 2004

Proton pumps participate in several aspects of endocytic protein trafficking. However, their involvement specifically in the GLUT4 pathway has been a matter of great controversy. Here, we report that incubation of 3T3-L1 adipocytes with specific inhibitors of V-type ATPase, concanamycin A and bafilomycin A1, inhibits insulin-regulated glucose transport and results in accumulation of GLUT4 in heavy, rapidly sedimenting intracellular membranes. Correspondingly, the amount of small responsive GLUT4 vesicles in concanamycin A- and bafilomycin A1-treated cells is decreased. We conclude that these drugs block translocation of GLUT4 in adipose cells by inhibiting formation of small insulin-responsive vesicles on donor intracellular membranes. At the same time, proton pump inhibitors do not affect insulin-dependent translocation of preexisting vesicles or GLUT4 sorting in recycling endosomes. On the contrary, wortmannin acutely inhibits insulin-dependent translocation of the preexisting vesicles but has no effect on vesicle formation.

cellugyrin; adipocytes; bafilomycin; concanamycin; wortmannin


Address for reprint requests and other correspondence: K. V. Kandror, Dept. of Biochemistry, Boston Univ. School of Medicine, K124D, 715 Albany St., Boston, MA 02118 (E-mail: kandror{at}biochem.bumc.bu.edu).






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