Insulin-like growth factor-I (IGF-I) and IGF binding proteins (IGFBPs) are trophic factors for cartilage and have been shown to be chondroprotective in animal models of osteoarthritis. IGFBP-5 is degraded in joint fluid and inhibition of IGFBP-5 degradation has been shown to enhance the trophic effects of IGF-I

Busby Jr., W. H.

Clemmons, D. R.

Kellner, D.

Lazerwith, S.

Radabaugh, M.

Rowland, M.

Sverdrup, F.

Yates, M.

Yocum, S. A.

SummaryInsulin-like growth factor-I (IGF-I) and IGF binding proteins (IGFBPs) are trophic factors for cartilage and have been shown to be chondroprotective in animal models of osteoarthritis (OA). IGFBP-5 is degraded in joint fluid and inhibition of IGFBP-5 degradation has been shown to enhance the trophic effects of IGF-I.ObjectiveTo determine the identity of IGFBP-5 protease activity in human OA joint fluid.MethodOA joint fluid was purified and the purified material was analyzed by IGFBP-5 zymography.ResultsBoth crude joint fluid and purified material contained a single band of proteolytic activity that cleaved IGFBP-5. Immunoblotting of joint fluid for complement 1s (C1s) showed a band that had the same Mr estimate, e.g., 88kDa. In gel tryptic digestion and subsequent peptide analysis by LC-MS/MS showed that the band contained human C1s. A panel of protease inhibitors was tested for their ability to inhibit IGFBP-5 cleavage by the purified protease. Three serine protease inhibitors, FUT175 and CP-143217 and CB-349547 had IC50's between 1 and 6μM. Two other serine protease inhibitors had intermediate activity (e.g., IC50's 20–40μM) and MMP inhibitors had no detectible activity at concentrations up to 300μM.ConclusionHuman OA fluid contains a serine protease that cleaves IGFBP-5. Zymography, immunoblotting and LC-MS/MS analysis indicate that C1s is the protease that accounts for this activity

Busby, W.H.

Yocum, S.A.

Clemmons, D.R.

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Complement 1s is the serine protease that cleaves IGFBP-5 in human osteoarthritic joint fluid 

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Complement 1s is the serine protease that cleaves IGFBP-5 in human osteoarthritic joint fluid

https://cdr.lib.unc.edu/downloads/bv73c7180

Complement 1s is the serine protease that cleaves IGFBP-5 in human osteoarthritic joint fluid

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