Characterization of metalloprotease cleavage products of human articular cartilage
| Autor: | D. A. Laska, Kevin L. Duffin, Eugene Y. Zhen, P.G. Mitchell, E.U. Sumer, Morten A. Karsdal, I.J. Brittain |
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| Rok vydání: | 2008 |
| Předmět: |
Adult
Cartilage Articular Lumican Molecular Sequence Data Immunology Type II collagen Chondrocyte Rheumatology Biglycan medicine Humans Immunology and Allergy Pharmacology (medical) Amino Acid Sequence Collagen Type II Aggrecan Cartilage oligomeric matrix protein Extracellular Matrix Proteins biology Chemistry Arthritis Cartilage Middle Aged ADAM Proteins Collagen type I alpha 1 medicine.anatomical_structure Biochemistry ADAMTS4 Protein Metalloproteases biology.protein Female Proteoglycans ADAMTS5 Protein Peptides Procollagen N-Endopeptidase Biomarkers |
| Zdroj: | Arthritis & Rheumatism. 58:2420-2431 |
| ISSN: | 1529-0131 0004-3591 |
| DOI: | 10.1002/art.23654 |
| Popis: | Objective To identify, characterize, and compare proteolysis peptide products generated by metalloprotease digests of human articular cartilage. Methods Human articular cartilage was digested by the addition of exogenous metalloproteases, including matrix metalloproteinases 2, 3, 8, 9, 12, and 13 and aggrecanases ADAMTS-4 and ADAMTS-5. Proteolyzed peptide products were identified by proteomics methods using mass spectrometry. Results Complete sequences of the peptides proteolyzed from human articular cartilage, including N- and C-termini and hydroxylated posttranslational modifications, were determined. A wide variety of peptides, originating from types I, II, and III collagen, biglycan, prolargin, fibromodulin, fibronectin, decorin, cartilage oligomeric matrix protein, cartilage intermediate-layer protein, megakaryocyte-stimulating factor, mimecan, aggrecan, and lumican, was analyzed following metalloprotease digestion. Release of peptides varied as a function of time, enzyme specificity, and abundance. Specific type II collagen peptide biomarkers, including those containing the three-quarter–length fragment cleavage site and those containing the domains for helical peptide of type II collagen and C-telopeptide of type II collagen, were observed after release by selected proteases. Conclusion The use of intact cartilage instead of purified protein substrates in the assay allowed for the identification of novel potential substrates and cleavage sites for individual enzymes under more physiologically relevant conditions. Characterization of these cartilage matrix peptides may help in the development of pharmacodynamic biomarkers of cartilage degradation, and also may contribute to an understanding of the bioactive peptides important in chondrocyte signaling. |
| Databáze: | OpenAIRE |
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