Functional duplication of ligand-binding domains within low-density lipoprotein receptor-related protein for interaction with receptor associated protein, alpha2-macroglobulin, factor IXa and factor VIII

Biochim Biophys Acta. 2007 Jun;1774(6):714-22. doi: 10.1016/j.bbapap.2007.04.003. Epub 2007 Apr 14.

Abstract

The low-density lipoprotein receptor-related protein (LRP) binds a range of proteins including receptor associated protein (RAP), activated alpha2-macroglobulin (alpha2M*), factor IXa (FIXa), and factor VIII (FVIII) light chain. The binding is mediated by the complement-type repeats, which are clustered in four distinct regions within LRP. Cluster II of 8 repeats (CR3-10) and cluster IV of 11 repeats (CR21-31) have been implicated in ligand-binding. Previous studies have aimed to identify the cluster II repeats involved in binding alpha2M* and RAP. We now evaluated the binding to RAP, alpha2M*, FIXa and FVIII light chain of triplicate repeat-fragments of not only clusters II but also of cluster IV. Employing surface plasmon resonance analysis, we found that most efficient ligand-binding was displayed by the repeats within region CR4-8 of cluster II and within region CR24-28 of cluster IV. Whereas the binding to RAP could be attributed to two consecutive repeats (CR5-6, CR26-27), combinations of three repeats showed most efficient binding to FIXa (CR6-8, CR26-28), FVIII light chain (CR5-7, CR6-8, CR24-26), and alpha2M* (CR4-6, CR24-26). The results imply that there is an internal functional duplication of complement-type repeats within LRP resulting in two clusters that bind the same ligands.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Binding Sites
  • Cell Line
  • Cricetinae
  • Factor IXa / metabolism*
  • Factor VIII / metabolism*
  • Glycosylation
  • Humans
  • LDL-Receptor Related Protein-Associated Protein / metabolism*
  • LDL-Receptor Related Proteins / chemistry
  • LDL-Receptor Related Proteins / genetics
  • LDL-Receptor Related Proteins / metabolism*
  • Ligands
  • Molecular Sequence Data
  • Multigene Family
  • Peptide Fragments / chemistry
  • Peptide Fragments / genetics
  • Peptide Fragments / metabolism
  • Protein Binding
  • Protein Structure, Tertiary
  • Sequence Alignment
  • Surface Plasmon Resonance
  • alpha-Macroglobulins / metabolism*

Substances

  • LDL-Receptor Related Protein-Associated Protein
  • LDL-Receptor Related Proteins
  • Ligands
  • Peptide Fragments
  • alpha-Macroglobulins
  • Factor VIII
  • Factor IXa