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

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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.

Original languageEnglish
Pages (from-to)834-838
Number of pages5
JournalCurrent Pharmaceutical Biotechnology
Volume12
Issue number5
DOIs
StatePublished - 1 Jan 2011

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Matrix Metalloproteinases
Fluorescence
Peptides
Enzymes
Fluorescence Resonance Energy Transfer
Fluorescein
Coloring Agents
Observation
cyanine dye 5

Keywords

  • FRET
  • Fluorescence
  • Labeled peptide
  • MMP-9

Cite this

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title = "Fluorescence detection of MMP-9. I. MMP-9 selectively cleaves Lys-Gly- Pro-Arg-Ser-Leu-Ser-Gly-Lys 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 F{\"o}rster 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 = "FRET, Fluorescence, Labeled peptide, MMP-9",
author = "Rafal Fudala and Ranjan, {Amalendu Prakash} and Anindita Mukerjee and Vishwanatha, {Jamboor K.} and Zygmunt Gryczynski and Julian Borejdo and Pabak Sarkar and Ignacy Gryczynski",
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T1 - Fluorescence detection of MMP-9. I. MMP-9 selectively cleaves Lys-Gly- Pro-Arg-Ser-Leu-Ser-Gly-Lys Peptide

AU - Fudala, Rafal

AU - Ranjan, Amalendu Prakash

AU - Mukerjee, Anindita

AU - Vishwanatha, Jamboor K.

AU - Gryczynski, Zygmunt

AU - Borejdo, Julian

AU - Sarkar, Pabak

AU - Gryczynski, Ignacy

PY - 2011/1/1

Y1 - 2011/1/1

N2 - 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.

AB - 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.

KW - FRET

KW - Fluorescence

KW - Labeled peptide

KW - MMP-9

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U2 - 10.2174/138920111795470967

DO - 10.2174/138920111795470967

M3 - Article

C2 - 21446907

AN - SCOPUS:79955612868

VL - 12

SP - 834

EP - 838

JO - Current Pharmaceutical Biotechnology

JF - Current Pharmaceutical Biotechnology

SN - 1389-2010

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