Fluorescence detection of MMP-9. I. MMP-9 selectively cleaves Lys-Gly-Pro-Arg-Ser-Leu-Ser-Gly-Lys peptide

Curr Pharm Biotechnol. 2011 May;12(5):834-8. doi: 10.2174/138920111795470967.

Abstract

MMP-9 enzyme recognizes a peptide sequence Lys-Gly-Pro-Arg-Ser-Leu-Ser-Gly-Lys and cleaves the peptide into two parts. We synthesized a dual fluorophore beacon consisting of 5-FAM and Cy5 dyes. The fluorescence emission of the fluorescein moiety is dramatically quenched by Cy5 molecule due to Förster Resonance Energy Transfer (FRET) and the fluorescence of Cy5 is strongly enhanced. Upon addition of MMP-9 enzyme, the fluorescence of 5-FAM intensifies and Cy5 decreases. The control MMP-2 enzyme does not cause any changes in either 5-FAM or Cy5 fluorescence. We believe that our observation will help in early detection of elevated MMP-9 levels under disease conditions.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Carbocyanines / chemistry
  • Fluorescence
  • Fluorescence Resonance Energy Transfer / methods*
  • Matrix Metalloproteinase 2 / analysis
  • Matrix Metalloproteinase 2 / metabolism
  • Matrix Metalloproteinase 9 / analysis*
  • Matrix Metalloproteinase 9 / metabolism
  • Oligopeptides / metabolism*
  • Peptide Fragments / analysis
  • Peptide Fragments / metabolism
  • Substrate Specificity

Substances

  • Carbocyanines
  • Oligopeptides
  • Peptide Fragments
  • cyanine dye 5
  • Matrix Metalloproteinase 2
  • Matrix Metalloproteinase 9