Cleavage site specificity of cathepsin K toward cartilage proteoglycans and protease complex formation.
Cathepsin K is a potent extracellular matrix-degrading protease that requires interactions with soluble glycosaminolycans for its collagenolytic activity in bone and cartilage. The major sources of glycosaminoglycans in cartilage are aggrecan aggregates. Therefore, we investigated whether cathepsin K activity is capable to hydrolyze aggrecan into fragments allowing the formation of glycosaminoglycan-cathepsin K complexes and determined the cleavage site specificity of cathepsin K toward the cartilage-resident link protein and aggrecan. The cleavage site specificity was compared with those of cathepsins S and L. All three cathepsins released glycosaminoglycans from native bovine cartilage at lysosomal pH and to a lesser degree at neutral extracellular pH. Cathepsin-predigested aggrecan complexes and cartilage provided suitable glycosaminoglycan fragments that allowed the formation of collagenolytically active cathepsin K complexes. A detailed analysis of the degradation of aggrecan aggregates revealed two cathepsin K cleavage sites in the link protein and several sites in aggrecan, including one site within the interglobular domain E1. In summary, these results demonstrate that cathepsin K is capable to degrade aggrecan complexes at specific cleavage sites and that cathepsin K activity alone is sufficient to self-provide the glycosaminoglycan fragments required for the formation of its collagenolytically active complex.
Hou
Wu-Shiun
WS
Department of Human Genetics, Mount Sinai School of Medicine, New York, NY 10029, USA.
Li
Zhenqiang
Z
Büttner
Frank H
FH
Bartnik
Eckart
E
Brömme
Dieter
D
D053674:Aggrecans; D000595:Amino Acid Sequence; D000818:Animals; D002356:Cartilage / Q000737:chemistry / Q000378:metabolism*; D056657:Cathepsin K; D002403:Cathepsins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D002417:Cattle; D016326:Extracellular Matrix Proteins; D006025:Glycosaminoglycans / Q000378:metabolism; D006801:Humans; D037181:Lectins, C-Type; D008969:Molecular Sequence Data; D009097:Multienzyme Complexes / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011506:Proteins / Q000737:chemistry / Q000378:metabolism; D011509:Proteoglycans / Q000737:chemistry / Q000378:metabolism*; D013379:Substrate Specificity; D000099249:Proteoglycan Link Protein
D053674:Aggrecans; D056657:Cathepsin K; D002403:Cathepsins; D016326:Extracellular Matrix Proteins; D006025:Glycosaminoglycans; D037181:Lectins, C-Type; D009097:Multienzyme Complexes; D011506:Proteins; D011509:Proteoglycans; D000099249:Proteoglycan Link Protein; C534130:CTSK protein, human