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Originally published In Press as doi:10.1074/jbc.M101691200 on March 26, 2001

J. Biol. Chem., Vol. 276, Issue 24, 21121-21128, June 15, 2001
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The Apolipoprotein E-dependent Low Density Lipoprotein Cholesteryl Ester Selective Uptake Pathway in Murine Adrenocortical Cells Involves Chondroitin Sulfate Proteoglycans and an alpha 2-Macroglobulin Receptor*

Snehasikta SwarnakarDagger , Jeanette Beers§, Dudley K. Strickland§, Salman Azhar, and David L. WilliamsDagger ||

From the Dagger  Department of Pharmacological Sciences, University Medical Center, State University of New York, Stony Brook, New York 11794, § American Red Cross, Holland Laboratory, Rockville, Maryland 20855, and  Geriatric Research, Education, and Clinical Center, Veterans Affairs, Palo Alto Health Care System, Palo Alto, California 94304

Cells acquire lipoprotein cholesterol by receptor-mediated endocytosis and selective uptake pathways. In the latter case, lipoprotein cholesteryl ester (CE) is transferred to the plasma membrane without endocytosis and degradation of the lipoprotein particle. Previous studies with Y1/E/tet/2/3 murine adrenocortical cells that were engineered to express apolipoprotein (apo) E demonstrated that apoE expression enhances low density lipoprotein (LDL) CE uptake by both selective and endocytic pathways. The present experiments test the hypothesis that apoE-dependent LDL CE selective uptake is mediated by scavenger receptor, class B, type I (SR-BI). Surprisingly, SR-BI expression was not detected in the Y1/E/tet/2/3 clone of Y1 adrenocortical cells, indicating the presence of a distinct apoE-dependent pathway for LDL CE selective uptake. ApoE-dependent LDL CE selective uptake in Y1/E/tet/2/3 cells was inhibited by receptor-associated protein and by activated alpha 2-macroglobulin (alpha 2M), suggesting the participation of the LDL receptor-related protein/alpha 2M receptor. Reagents that inhibited proteoglycan synthesis or removed cell surface chondroitin sulfate proteoglycan completely blocked apoE-dependent LDL CE selective uptake. None of these reagents inhibited SR-BI-mediated LDL CE selective uptake in the Y1-BS1 clone of Y1 cells in which LDL CE selective uptake is mediated by SR-BI. We conclude that LDL CE selective uptake in adrenocortical cells occurs via SR-BI-independent and SR-BI-dependent pathways. The SR-BI-independent pathway is an apoE-dependent process that involves both chondroitin sulfate proteoglycans and an alpha 2M receptor.

* This work was supported by National Institutes of Health Grants HL 58012, HL 32868, HL 50784, and HL 54710 and by the Office of Research and Development, Medical Research Service, Department of Veterans Affairs.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: Dept. of Pharmacological Sciences, University Medical Center, State University of New York, Stony Brook, NY 11794. Tel.: 516-444-3083; Fax: 516-444-3218; E-mail: dave@pharm.som.sunysb.edu.

Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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