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J. Biol. Chem., Vol. 277, Issue 24, 21149-21157, June 14, 2002
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From the The high density lipoprotein receptor, scavenger
receptor class B type I (SR-BI), recognizes lipid-bound apolipoprotein
A-I (apoA-I) and other apolipoproteins. Here, we have used large scale cultures of apoE-expressing cells to purify apoE and prepare apoE containing reconstituted discoidal
1-palmitoyl-2-oleoyl-L-phosphatidylcholine (POPC)-apoE particles. These particles have been used to examine their
binding to wild-type and mutant forms of SR-BI expressed in transfected
ldlA-7 cells. Specific binding to SR-BI was determined by subtracting
from the total binding, nonspecific values measured using either
control untransfected ldlA-7 cells or by inhibiting SR-BI-mediated
binding with a high titer antireceptor-blocking antibody. POPC-apoE
particles generated using apoE2, apoE3, apoE4, or the
carboxyl-terminally truncated forms apoE165, apoE202, apoE229, and apoE259 all bound tightly to wild-type SR-BI with similar affinities (Kd = 35-45 µg/ml). Binding was
nearly abolished in a cell line expressing the ldlA (Q402R/Q418R)
double mutant form of SR-BI that is unable to bind native high
density lipoprotein but binds low density lipoprotein normally. The
findings establish that apoE is a ligand for SR-BI and that the
receptor binding domain is located in the amino-terminal 1-165-region
of the protein. SR-BI-apoE interactions may contribute to cholesterol
homeostasis in tissues and cells expressing SR-BI that are accessible
to apoE-containing lipoproteins.
Reconstituted Discoidal ApoE-Phospholipid Particles Are Ligands
for the Scavenger Receptor BI
THE AMINO-TERMINAL 1-165 DOMAIN OF ApoE SUFFICES FOR RECEPTOR
BINDING*
,,,¶
Section of Molecular Genetics, Whitaker
Cardiovascular Institute, Departments of Medicine and Biochemistry,
Boston University School of Medicine, Boston, Massachusetts 02118 and
the § Biology Department, Massachusetts Institute of
Technology, Cambridge, Massachusetts 02139
*
This work was supported by National Institutes of Health
Grants HL48739 and HL68216 (to V. Z.) and HL41484 and HL52212 (to M. K.) and the Alzheimer's Association Grant IIRG-002220 (to V. Z.).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.
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