Utility of combining MMP-9 enzyme-linked immunosorbent assay and MMP-9 activity assay data to monitor plasma enzyme specific activity.
The aim of this study was to combine matrix metalloproteinase-9 (MMP-9) protein (enzyme-linked immunosorbent assay [ELISA]) and MMP-9 activity (fluorescence resonance energy transfer [FRET] assay) data to generate units of specific activity in endogenous and p-aminophenylmercuric acetate (APMA)-activated lithium heparin plasma. The results indicate that specific activity is constant in APMA-activated plasma (mean value=1359.4 pmol/min/microg) and approximately 12% plasma MMP-9 is endogenously active. Exogenous tissue inhibitor of metalloproteinase-1 (TIMP-1) has a greater inhibitory effect on endogenously active MMP-9 than on APMA-activated MMP-9. In conclusion, specific activity can be used as a tool to monitor MMP-9 inhibition. APMA activation affects natural enzyme inhibition, possibly by chemical modification of the C-terminal portion of the enzyme containing the TIMP-1 binding site.
Grierson
C
C
Translational Medicine Research Collaboration (TMRC), University of Dundee, Dundee, Angus DD14HN, Scotland, UK. c.grierson@dundee.ac.uk
Miller
D
D
LaPan
P
P
Brady
J
J
D001665:Binding Sites; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D031541:Fluorescence Resonance Energy Transfer; D020780:Matrix Metalloproteinase 9 / Q000097:blood*; D010662:Phenylmercuric Acetate / Q000031:analogs & derivatives / Q000737:chemistry; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000737:chemistry / Q000494:pharmacology
D019715:Tissue Inhibitor of Metalloproteinase-1; C010024:4-aminophenylmercuriacetate; D020780:Matrix Metalloproteinase 9; D010662:Phenylmercuric Acetate