Effect of arginine 172 on the binding of apolipoprotein E to the low density lipoprotein receptor.
The region of apolipoprotein E (apoE) that interacts directly with the low density lipoprotein (LDL) receptor lies in the vicinity of residues 136-150, where lysine and arginine residues are crucial for full binding activity. However, defective binding of carboxyl-terminal truncations of apoE3 has suggested that residues in the vicinity of 170-183 are also important. To characterize and define the role of this region in LDL receptor binding, we created either mutants of apoE in which this region was deleted or in which arginine residues within this region were sequentially changed to alanine. Deletion of residues 167-185 reduced binding activity (15% of apoE3), and elimination of arginines at positions 167, 172, 178, and 180 revealed that only position 172 affected binding activity (2% of apoE3). Substitution of lysine for Arg(172) reduced binding activity to 6%, indicating a specific requirement for arginine at this position. The higher binding activity of the Delta167-185 mutant relative to the Arg(172) mutant (15% versus 2%) is explained by the fact that arginine residues at positions 189 and 191 are shifted in the deletion mutant into positions equivalent to 170 and 172 in the intact protein. Mutation of these residues and modeling the region around these residues suggested that the influence of Arg(172) on receptor binding activity may be determined by its orientation at a lipid surface. Thus, the association of apoE with phospholipids allows Arg(172) to interact directly with the LDL receptor or with other residues in apoE to promote its receptor-active conformation.
Morrow
J A
JA
Gladstone Institute of Cardiovascular Disease, University of California, San Francisco, California 94141-9100, USA.
Arnold
K S
KS
Dong
J
J
Balestra
M E
ME
Innerarity
T L
TL
Weisgraber
K H
KH
D000595:Amino Acid Sequence; D001057:Apolipoproteins E / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D001120:Arginine / Q000378:metabolism*; D001483:Base Sequence; D018076:DNA, Complementary; D006801:Humans; D008969:Molecular Sequence Data; D017354:Point Mutation; D011485:Protein Binding; D011973:Receptors, LDL / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid
D001057:Apolipoproteins E; D018076:DNA, Complementary; D011973:Receptors, LDL; D001120:Arginine