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.
Keywords: MMP-9, fluorescence, FRET, labeled peptide
Current Pharmaceutical Biotechnology
Title: Fluorescence Detection of MMP-9. I. MMP-9 Selectively Cleaves Lys-Gly-Pro-Arg-Ser-Leu-Ser-Gly-Lys Peptide
Volume: 12 Issue: 5
Author(s): Rafal Fudala, Amalendu P. Ranjan, Anindita Mukerjee, Jamboor K. Vishwanatha, Zygmunt Gryczynski, Julian Borejdo, Pabak Sarkar and Ignacy Gryczynski
Affiliation:
Keywords: MMP-9, fluorescence, FRET, labeled peptide
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.
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Fudala Rafal, P. Ranjan Amalendu, Mukerjee Anindita, K. Vishwanatha Jamboor, Gryczynski Zygmunt, Borejdo Julian, Sarkar Pabak and Gryczynski Ignacy, Fluorescence Detection of MMP-9. I. MMP-9 Selectively Cleaves Lys-Gly-Pro-Arg-Ser-Leu-Ser-Gly-Lys Peptide, Current Pharmaceutical Biotechnology 2011; 12 (5) . https://dx.doi.org/10.2174/138920111795470967
DOI https://dx.doi.org/10.2174/138920111795470967 |
Print ISSN 1389-2010 |
Publisher Name Bentham Science Publisher |
Online ISSN 1873-4316 |
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