Utility of combining MMP-9 enzyme-linked immunosorbent assay and MMP-9 activity assay data to monitor plasma enzyme specific activity

Anal Biochem. 2010 Sep 15;404(2):232-4. doi: 10.1016/j.ab.2010.05.020. Epub 2010 May 25.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Binding Sites
  • Enzyme-Linked Immunosorbent Assay / methods*
  • Fluorescence Resonance Energy Transfer
  • Matrix Metalloproteinase 9 / blood*
  • Phenylmercuric Acetate / analogs & derivatives
  • Phenylmercuric Acetate / chemistry
  • Tissue Inhibitor of Metalloproteinase-1 / chemistry
  • Tissue Inhibitor of Metalloproteinase-1 / pharmacology

Substances

  • Tissue Inhibitor of Metalloproteinase-1
  • 4-aminophenylmercuriacetate
  • Matrix Metalloproteinase 9
  • Phenylmercuric Acetate