Identification and quantification of degradome components in human synovial fluid reveals an increased proteolytic activity in knee osteoarthritis patients vs controls

Proteomics. 2023 Aug;23(15):e2300040. doi: 10.1002/pmic.202300040. Epub 2023 May 24.


Synovial fluid (SF) may contain cleavage products of proteolytic activities. Our aim was to characterize the degradome through analysis of proteolytic activity and differential abundance of these components in a peptidomic analysis of SF in knee osteoarthritis (OA) patients versus controls (n = 23). SF samples from end-stage knee osteoarthritis patients undergoing total knee replacement surgery and controls, that is, deceased donors without known knee disease were previously run using liquid chromatography mass spectrometry (LC-MS). This data was used to perform new database searches generating results for non-tryptic and semi-tryptic peptides for studies of degradomics in OA. We used linear mixed models to estimate differences in peptide-level expression between the two groups. Known proteolytic events (from the MEROPS peptidase database) were mapped to the dataset, allowing the identification of potential proteases and which substrates they cleave. We also developed a peptide-centric R tool, proteasy, which facilitates analyses that involve retrieval and mapping of proteolytic events. We identified 429 differentially abundant peptides. We found that the increased abundance of cleaved APOA1 peptides is likely a consequence of enzymatic degradation by metalloproteinases and chymase. We identified metalloproteinase, chymase, and cathepsins as the main proteolytic actors. The analysis indicated increased activity of these proteases irrespective of their abundance.

Keywords: degradomics; osteoarthritis; peptidomics; proteomics; synovial fluid.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chymases / analysis
  • Chymases / metabolism
  • Humans
  • Osteoarthritis, Knee* / metabolism
  • Peptide Hydrolases / analysis
  • Peptides / analysis
  • Synovial Fluid / chemistry
  • Synovial Fluid / metabolism


  • Chymases
  • Peptide Hydrolases
  • Peptides