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J. Biol. Chem., Vol. 275, Issue 45, 35264-35275, November 10, 2000
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From the Lateral association between different
transmembrane glycoproteins can serve to modulate integrin function.
Here we characterize a physical association between the integrins
The Integrins
3
1 and
61 Physically and Functionally Associate
with CD36 in Human Melanoma Cells
REQUIREMENT FOR THE EXTRACELLULAR DOMAIN OF CD36*
§,
,
Cancer Research Unit and Department
of Microbiology, Faculty of Medicine and Health Sciences, University of
Newcastle, New South Wales 2308, Australia, the ** Department of
Respiratory Medicine, John Hunter Hospital, Newcastle, New South Wales
2305, Australia, and the ¶ Richard Dimbleby Department of Cancer
Research/Imperial Cancer Research Fund, Rayne Institute, St Thomas'
Hospital, London SE1 7EH, United Kingdom
3
1 and
61 and CD36 on the surface of melanoma
cells and show that ectopic expression of CD36 by CD36-negative MV3
melanoma cells increases their haptotactic migration on extracellular
matrix components. The association was demonstrated by
co-immunoprecipitation, reimmunoprecipitation, and immunoblotting of
surface-labeled cells lysed in Brij 96 detergent. Confocal microscopy
illustrated the co-association of 3 and CD36 in cell
membrane projections and ruffles. A requirement for the extracellular
domain of CD36 in this association was shown by co-immunoprecipitation
experiments using surface-labeled MV3 melanoma or COS-7 cells that had
been transiently transfected with chimeric constructs between CD36 and
intercellular adhesion molecule 1 (ICAM-1) or with a truncation mutant
of CD36. CD36 is known to engage in signal transduction and to localize
to membrane microdomains or rafts in several cell types. Toward a
mechanistic explanation for the functional effects of CD36 expression,
we demonstrate that in fractionated Triton X-100 lysates of the MV3
cells stably transfected with CD36, CD36 was greatly enriched with the
detergent-insoluble fractions that represent plasma membrane rafts.
Significantly, when these fractionated lysates were reprobed for
endogenous 1 integrin, it was found that a 4-fold
increase in the proportion of the mature protein was contained within
the detergent-insoluble fractions when extracted from the
CD36-transfected cells compared with MV3 cells transfected with vector
only. These results suggest that in melanoma cells CD36 expression may
induce the sequestration of certain integrins into membrane
microdomains and promote cell migration.
*
This work was supported by the National Health & Medical
Research Council (Australia).The costs of publication of this
article were defrayed in part by the
payment of page charges. The article must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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