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J. Biol. Chem., Vol. 277, Issue 35, 32187-32194, August 30, 2002
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From the ¶ Institute for Medical Biochemistry and Molecular
Biology, Department of Molecular Cell Biology, University Hospital
Eppendorf, D-20246 Hamburg, Germany and the Annexins are Ca2+- and
phospholipid-binding proteins that are widely expressed in
mammalian tissues and that bind to different cellular membranes.
In recent years its role in membrane traffic has emerged as one of its
predominant functions, but the regulation of its intracellular
distribution still remains unclear. We demonstrated that annexin 6 translocates to the late endocytic compartment in low density
lipoprotein-loaded CHO cells. This prompted us to investigate whether
cholesterol, one of the major constituents of low density lipoprotein,
could influence the membrane binding affinity and intracellular
distribution of annexin 6. Treatment of crude membranes or early and
late endosomal fractions with digitonin, a cholesterol-sequestering
agent, displayed a strong reduction in the binding affinity of a novel
EDTA-resistant and cholesterol-sensitive pool of annexin 6 proteins. In
addition, U18666A-induced accumulation of cholesterol in the late
endosomal compartment resulted in a significant increase of annexin 6 in these vesicles in vivo. This translocation/recruitment
correlates with an increased membrane binding affinity of GST-annexin 6 to late endosomes of U18666A-treated cells in vitro. In
conclusion, the present study shows that changes in the intracellular
distribution and concentration of cholesterol in different subcellular
compartments participate in the reorganization of intracellular pools
of Ca2+-dependent and -independent annexin 6.
Cholesterol Modulates the Membrane Binding and
Intracellular Distribution of Annexin 6*
§,, and
Departament
de Biología Cellular, Institut d'Investigacions
Biomèdiques August Pi i Sunyer (IDIBAPS), Facultat de Medicina,
Universitat de Barcelona, 0836 Barcelona, Spain
*
This work was supported by the Deutsche
Forschungsgemeinschaft (DFG, Ja 421/3-1 and Be 829/5-1) and Grants from
Ministerio de Ciencia y Tecnología (PM99-0166), Acciones
Integradas (HA98-0007), and Generalitat de Catalunya (BE2000).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.
To whom correspondence should be addressed: Inst. for Medical
Biochemistry and Molecular Biology, Dept. of Molecular Cell Biology,
University Hospital Hamburg-Eppendorf, Martinistr. 52, D-20246 Hamburg,
Germany. Tel.: 49-40-42803-4745; Fax: 49-40-42803-4592; E-mail:
grewal@uke.uni-hamburg.de.
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