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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 publication

DOI

10.1042/BJ20112159

Type

Journal article

Journal

Biochem J

Publication Date

01/04/2012

Volume

443

Pages

307 - 315

Keywords

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