Pentosan polysulfate increases affinity between ADAMTS-5 and TIMP-3 through formation of an electrostatically driven trimolecular complex

Linda Troeberg, Barbara Mulloy, Peter Ghosh, Meng-Huee Lee, Gillian Murphy, Hideaki Nagase

Research output: Contribution to journalArticlepeer-review

23 Citations (Scopus)

Abstract

The semi-synthetic sulfated polysaccharide PPS (pentosan polysulfate) increases affinity between the aggrecan-degrading ADAMTSs (adamalysins with thrombospondin motifs) and their endogenous inhibitor, TIMP (tissue inhibitor of metalloproteinases)-3. In the present study we demonstrate that PPS mediates the formation of a high-affinity trimolecular complex with ADAMTS-5 and TIMP-3. A TIMP-3 mutant that lacks extracellular-matrix-binding ability was insensitive to this affinity increase, and truncated forms of ADAMTS-5 that lack the Sp (spacer) domain had reduced PPS-binding ability and sensitivity to the affinity increase. PPS molecules composed of 11 or more saccharide units were 100-fold more effective than those of eight saccharide units, indicating the involvement of extended or multiple protein-interaction sites. The formation of a high-affinity trimolecular complex was completely abolished in the presence of 0.4 M NaCl. These results suggest that PPS enhances the affinity between ADAMTS-5 and TIMP-3 by forming electrostatically driven trimolecular complexes under physiological conditions.

Original languageEnglish
Pages (from-to)307-315
Number of pages9
JournalBiochemical Journal
Volume443
Issue number1
DOIs
Publication statusPublished - 1 Apr 2012

Keywords

  • ADAM Proteins/biosynthesis
  • ADAMTS5 Protein
  • Amino Acid Substitution
  • Chromatography, Gel
  • HEK293 Cells
  • Humans
  • Pentosan Sulfuric Polyester/isolation & purification
  • Protein Binding
  • Protein Structure, Tertiary
  • Recombinant Proteins/biosynthesis
  • Sequence Deletion
  • Sodium Chloride/chemistry
  • Tissue Inhibitor of Metalloproteinase-3/biosynthesis

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