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Vol. 10, Issue 5, 1581-1594, May 1999
Department of Biochemistry, Boston University School of Medicine,
Boston, Massachusetts 02118
Differentiating 3T3-L1 cells exhibit a dramatic increase in the
rate of insulin-stimulated glucose transport during their conversion
from proliferating fibroblasts to nonproliferating adipocytes. On day 3 of 3T3-L1 cell differentiation, basal glucose transport and cell
surface transferrin binding are markedly diminished. This occurs
concomitant with the formation of a distinct insulin-responsive vesicular pool of intracellular glucose transporter 1 (GLUT1) and
transferrin receptors as assessed by sucrose velocity gradients. The
intracellular distribution of the insulin-responsive
aminopeptidase is first readily detectable on day 3, and its gradient profile and response to insulin at this time are
identical to that of GLUT1. With further time of differentiation, GLUT4
is expressed and targeted to the same insulin-responsive vesicles as
the other three proteins. Our data are consistent with the notion that
a distinct insulin-sensitive vesicular cargo compartment forms early during fat call differentiation and its formation precedes GLUT4 expression. The development of this compartment may result from the
differentiation-dependent inhibition of constitutive GLUT1 and
transferrin receptor trafficking such that there is a large increase
in, or the new formation of, a population of postendosomal, insulin-responsive vesicles.
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