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

Yamamoto, K; Owen, K; Parker, A E; Scilabra, S D; Dudhia, J; Strickland, D K; Troeberg, L; Nagase, H

Abstract

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.

Citation

Yamamoto, K., Owen, K., Parker, A. E., Scilabra, S. D., Dudhia, J., Strickland, D. K., …Nagase, H. (2014). 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. Journal of Biological Chemistry, 289(10), 6462-6474. https://doi.org/10.1074/jbc.M113.545376

• Journal Article Type: Article
• Acceptance Date: Jan 24, 2014
• Publication Date: Mar 7, 2014
• Deposit Date: Nov 11, 2014
• Publicly Available Date: Jul 5, 2018
• Journal: JOURNAL OF BIOLOGICAL CHEMISTRY
• Print ISSN: 0021-9258
• Publisher: American Society for Biochemistry and Molecular Biology
• Peer Reviewed: Peer Reviewed
• Volume: 289
• Issue: 10
• Pages: 6462-6474
• DOI: https://doi.org/10.1074/jbc.M113.545376
• Public URL: https://rvc-repository.worktribe.com/output/1405875