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J Biol Chem, Vol. 273, Issue 14, 8025-8032, April 3, 1998
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From the The previous characterization of an
abundant population of non-adrenergic imidazoline-I2
binding sites in adipocytes and the recent demonstration of the
interplay between these binding sites and amine oxidases led us to
analyze the amine oxidase activity in membranes from isolated rat
adipocytes. Adipocyte membranes had substantial levels of
semicarbazide-sensitive amine oxidase (SSAO). SSAO activity and
immunoreactive SSAO protein were maximal in plasma membranes, and they
were also detectable in intracellular membranes. Vesicle
immunoisolation analysis indicated that GLUT4-containing vesicles from
rat adipocytes contain substantial levels of SSAO activity and
immunoreactive SSAO protein. Immunotitration of intracellular GLUT4
vesicles indicated that GLUT4 and SSAO colocalize in an endosomal
compartment in rat adipocytes. SSAO activity was also found in GLUT4
vesicles from 3T3-L1 adipocytes and rat skeletal muscle.
Benzylamine, a substrate of SSAO activity, caused a marked stimulation
of glucose transport in isolated rat adipocytes in the presence
of very low vanadate concentrations that by themselves were ineffective
in exerting insulin-like effects. This synergistic effect of
benzylamine and vanadate on glucose transport was totally abolished in
the presence of semicarbazide, a specific inhibitor of SSAO.
Subcellular membrane fractionation revealed that the combination of
benzylamine and vanadate caused a recruitment of GLUT4 to the plasma
membrane of adipose cells. The stimulatory effects of benzylamine and
vanadate on glucose transport were blocked by catalase, suggesting that
hydrogen peroxide production coupled to SSAO activity plays a crucial
regulatory role. Based on these results we propose that SSAO activity
might contribute through hydrogen peroxide production to the in
vivo regulation of GLUT4 trafficking in adipose cells.
Departament de Bioquímica
i Biologia Molecular, Facultat de Biologia, Universitat de
Barcelona, Avinguda Diagonal 645, 08028 Barcelona, Spain,
INSERM U 317, Institut Louis Bugnard, Université Paul
Sabatier, Centre Hospitalaire Universitaire Rangueil, 31403 Toulouse Cedex 4, France, Departament de Bioquímica i
Biologia Molecular, Facultat de Medicina, Universitat Autònoma de
Barcelona, 08290 Barcelona, Spain
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