Cathepsin g Degrades Both Glycosylated and Unglycosylated Regions of Lubricin, a Synovial Mucin
| Autor: | Shan Huang, Thomas Eisler, Sebastian Kalamajski, Lena Björkman, Kristina A. Thomsson, Niclas G. Karlsson, Ola Rolfson, Tannin A. Schmidt, Roman Krawetz, Chunsheng Jin, Sally Alweddi, Gregory D. Jay, André Struglics |
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| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
medicine.medical_treatment lcsh:Medicine Cathepsin G Biochemistry Article law.invention 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Medical research Western blot Rheumatology law medicine Synovial fluid lcsh:Science Multidisciplinary Protease medicine.diagnostic_test lcsh:R Mucin Molecular biology Glycopeptide 030104 developmental biology chemistry Protein Fragment Recombinant DNA lcsh:Q Structural biology 030217 neurology & neurosurgery Biomarkers |
| Zdroj: | Scientific Reports Scientific Reports, Vol 10, Iss 1, Pp 1-12 (2020) |
| ISSN: | 2045-2322 |
| Popis: | Lubricin (PRG4) is a mucin type protein that plays an important role in maintaining normal joint function by providing lubrication and chondroprotection. Improper lubricin modification and degradation has been observed in idiopathic osteoarthritis (OA), while the detailed mechanism still remains unknown. We hypothesized that the protease cathepsin G (CG) may participate in degrading lubricin in synovial fluid (SF). The presence of endogenous CG in SF was confirmed in 16 patients with knee OA. Recombinant human lubricin (rhPRG4) and native lubricin purified from the SF of patients were incubated with exogenous CG and lubricin degradation was monitored using western blot, staining by Coomassie or Periodic Acid-Schiff base in gels, and with proteomics. Full length lubricin (∼300 kDa), was efficiently digested with CG generating a 25-kDa protein fragment, originating from the densely glycosylated mucin domain (∼250 kDa). The 25-kDa fragment was present in the SF from OA patients, and the amount was increased after incubation with CG. A CG digest of rhPRG4 revealed 135 peptides and 72 glycopeptides, and confirmed that the protease could cleave in all domains of lubricin, including the mucin domain. Our results suggest that synovial CG may take part in the degradation of lubricin, which could affect the pathological decrease of the lubrication in degenerative joint disease. |
| Databáze: | OpenAIRE |
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