Studies of aggrecan proteolysis in human joints have implicated both the aggrecanase [ADAMTS, a disintegrin-like and metalloprotease (reprolysin-type) with thrombospondin type 1 motif] and matrix metalloproteinase (MMP) families. We have analysed the aggrecan core protein species present in vivo in both articular cartilage and synovial fluids from normal, acutely injured and osteoarthritic joints. Normal cartilage contains at least seven major G1 domain (the N-terminal globular domain of aggrecan)-bearing species, of which three (full-length core, G1-NITEGE373 and G1-VDIPEN341) have been identified. The C-terminals of the others are unknown but digestion of fetal human aggrecan with MMP-3 and crude aggrecanase suggests that they are products of MMP-like activity in vivo. Normal synovial fluids contain at least 10 species, of which nine result from ADAMTS-dependent cleavage, and this cleavage occurs at all of the five known aggrecanase sites. Aggrecan fragments in the cartilage and synovial fluids of acutely injured joints are generally similar to normal, but all contain a markedly increased ratio of G1-NITEGE to G1-VDIPEN. Aggrecan from the cartilage of late-stage osteoarthritis patients is remarkably similar to normal, whereas the synovial fluid aggrecan is more fragmented than that from normal or injured knees. The analyses suggest that the role of the ADAMTS and these MMP-like activities in human cartilage are distinctly different. Excessive ADAMTS activity in vivo is destructive to cartilage matrix, since the bulk of the glycosaminoglycan (GAG)-bearing products are released from the tissue into the synovial fluid following cleavage of the Glu373–Ala374 bond. In contrast, the MMP-like activity appears to be essentially non-destructive, since much of the GAG-bearing product is retained in the tissue following cleavages that are in the more C-terminal regions of the molecule.
Abbreviations used: ADAMTS, a disintegrin-like and metalloprotease (reprolysin-type) with thrombospondin type 1 motif; MMP, matrix metalloproteinase; G1 domain, the N-terminal globular domain of aggrecan; G2 domain, the second N-terminal globular domain of aggrecan; G3 domain, the C-terminal globular domain of aggrecan; KS, keratan sulphate; CS, chondroitin sulphate; OA, osteoarthritis; AEBSF, 4-(2-aminoethyl)benzenesulphonyl fluoride; GAG, glycosaminoglycan; DE52, diaminoethylcellulose anion exchanger; ACL, anterior cruciate ligament.
- The Biochemical Society, London ©2001