Title
Highly increased levels of active stromelysin in rheumatoid synovial fluid determined by a selective fluorogenic assay
Author
Gaubius Instituut TNO
Beekman, B.
van El, B.
Drijfhout, J.W.
Ronday, H.K.
TeKoppele, J.M.
Publication year
1997
Abstract
Stromelysin-1 (MMP-3) is an important member of the matrix metalloproteinase family. In joint-degrading diseases like arthritis, elevated levels of MMP-3 protein are detected in synovial fluid using immunological methods. However, these methods do not discriminate between active and inactive enzyme. In the present study, a specific stromelysin activity assay was developed using the selective fluorogenic substrate TNO003 (Dabcyl-Gaba-Arg-Pro-Lys-Pro-Val-Glu · Nva-Trp-Arg-Glu-(EDANS)-Ala-Lys-NH2, · = cleavage site). For its use in biological media, cleavage of TNO003 by enzymes other than stromelysin was effectively blocked by a proteinase inhibitor cocktail. Spiking of MMP-3 to synovial fluid resulted in an MMP-3 concentration-dependent linear increase in activity. The measured MMP-3 activity was not affected by the addition of MMP-13, even in a 5-fold excess over MMP-3. Synovial fluid from rheumatoid arthritis patients demonstrated 100-fold higher levels of active stromelysin than control synovial fluids. Chemicals/CAS: Biological Markers; Matrix Metalloproteinase 3, EC 3.4.24.17
Subject
Biomedical Research
Fluorogenic substrate
MMP-3
Rheumatoid arthritis
Stromelysin
Synovial fluid
Stromelysin
Controlled study
Enzyme activity
Enzyme assay
Human tissue
Priority journal
Rheumatoid arthritis
Synovial fluid
Technique
Arthritis, Rheumatoid
Biological Assay
Biological Markers
Fluorescence
Humans
Matrix Metalloproteinase 3
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DOI
https://doi.org/10.1016/s0014-5793(97)01371-9
TNO identifier
234219
ISSN
0014-5793
Source
FEBS Letters, 418 (418), 305-309
Document type
article