Decoding of lipoprotein-receptor interactions: properties of ligand binding modules governing interactions with apolipoprotein E

Biochemistry. 2010 Feb 16;49(6):1207-16. doi: 10.1021/bi9017208.

Abstract

Clusters of complement-type ligand binding repeats in the LDL receptor family are thought to mediate the interactions between these receptors and their various ligands. Apolipoprotein E, a key ligand for cholesterol homeostasis, has been shown to interact with LDLR, LRP, and VLDLR, through these clusters. LDLR and VLDLR each contain a single ligand binding repeat cluster, whereas LRP contains three large clusters of ligand binding repeats, each with ligand binding functions. We show that within sLRP3 the three-repeat subcluster CR16-18 recapitulated ligand binding to the isolated receptor binding portion of ApoE (residues 130-149). Binding experiments with LA3-5 of LDLR and CR16-18 showed that a conserved W25/D30 pair appears to be critical for high-affinity binding to ApoE(130-149). The triple repeat LA3-5 showed the expected interaction with ApoE(1-191).DMPC, but surprisingly CR16-18 did not interact with this form of ApoE. To understand these differences in ApoE binding affinity, we introduced mutations of conserved residues from LA5 into CR18 and produced a CR16-18 variant capable of binding ApoE(1-191).DMPC. This change cannot fully be accounted for by the interaction with the proposed ApoE receptor binding region; therefore, we speculate that LA5 is recognizing a distinct epitope on ApoE that may only exist in the lipid-bound form. The combination of avidity effects with this distinct recognition process likely governs the ApoE-LDL receptor interaction.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Amino Acid Sequence
  • Apolipoproteins E / metabolism
  • Humans
  • Ligands
  • Low Density Lipoprotein Receptor-Related Protein-1 / chemistry
  • Low Density Lipoprotein Receptor-Related Protein-1 / metabolism*
  • Molecular Sequence Data
  • Protein Binding
  • Protein Structure, Tertiary
  • Receptors, LDL / chemistry
  • Receptors, LDL / metabolism*
  • Sequence Alignment

Substances

  • Apolipoproteins E
  • Ligands
  • Low Density Lipoprotein Receptor-Related Protein-1
  • Receptors, LDL
  • VLDL receptor