Low density lipoprotein receptor-related protein 1 (LRP1)-mediated endocytic clearance of a disintegrin and metalloproteinase with thrombospondin motifs-4 (ADAMTS-4): functional differences of non-catalytic domains of ADAMTS-4 and ADAMTS-5 in LRP1 binding

Kazuhiro Yamamoto, Kathryn Owen, Andrew E Parker, Simone D Scilabra, Jayesh Dudhia, Dudley K Strickland, Linda Troeberg, Hideaki Nagase

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Degradation of the cartilage proteoglycan aggrecan is an early event in the development of osteoarthritis, and a disintegrin and metalloproteinase with thrombospondin motifs-4 (ADAMTS-4) and ADAMTS-5 are considered to be the major aggrecan-degrading enzymes. We have recently found that ADAMTS-5 is rapidly endocytosed via low density lipoprotein receptor-related protein 1 (LRP1) and degraded by chondrocytes. Here we report that this regulatory mechanism also applies to ADAMTS-4, although its rate of endocytosis is slower than that of ADAMTS-5. Domain deletion mutagenesis of ADAMTS-4 identified that the cysteine-rich and spacer domains are responsible for binding to LRP1, whereas the thrombospondin 1 and spacer domains are responsible in ADAMTS-5. The estimated t½ value of ADAMTS-4 endocytosis was about 220 min, whereas that of ADAMTS-5 was 100 min. The difference in half-lives between the two enzymes is explained by the 13-fold lower affinity of ADAMTS-4 for LRP1 compared with that of ADAMTS-5. Studies using soluble ligand binding clusters of LRP1 showed that ADAMTS-4 binds to clusters II and IV with similar KD,app values of 98 and 73 nm, respectively, whereas ADAMTS-5 binds to cluster II, III, and IV with KD,app values of 3.5, 41, and 9 nm, respectively. Thus, ADAMTS-5 competitively inhibits ADAMTS-4 endocytosis but not vice versa. This study highlights that the affinity between a ligand and LRP1 dictates the rate of internalization and suggests that LRP1 is a major traffic controller of the two aggrecanases, especially under inflammatory conditions, where the protein levels of ADAMTS-4 increase, but those of ADAMTS-5 do not.

Original languageEnglish
Pages (from-to)6462-6474
Number of pages13
JournalThe Journal of Biological Chemistry
Issue number10
Publication statusPublished - 7 Mar 2014


  • ADAM Proteins/chemistry
  • ADAMTS4 Protein
  • ADAMTS5 Protein
  • Animals
  • Cartilage, Articular/metabolism
  • Catalytic Domain/genetics
  • Cells, Cultured
  • Endocytosis
  • Half-Life
  • Humans
  • Low Density Lipoprotein Receptor-Related Protein-1/metabolism
  • Osteoarthritis/metabolism
  • Procollagen N-Endopeptidase/chemistry
  • Protein Binding
  • Sequence Deletion
  • Swine

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