Fluorescence detection of MMP-9. II. ratiometric FRET-based sensing with dually labeled specific peptide

Research output: Contribution to journalArticleResearchpeer-review

9 Citations (Scopus)

Abstract

In our previous paper we showed that the MMP-9 enzyme recognizes a specific peptide sequence, Lys-Gly-Pro-Arg-Ser-Leu-Ser-Gly-Lys, and cleaves the peptide into two parts [1]. In this study, the peptide is labeled with two dyes, carboxyfluorescein (5-FAM) and Cy5. A highly efficient energy transfer of over 80% results in a dominant emission of Cy5 at ~670 nm with an excitation of 470 nm. Severance of the peptide by the MMP-9 enzyme eliminates Förster Resonance Energy Transfer (FRET) and strongly increases the fluorescence of the 5-FAM dye. In this manuscript we describe the strategy for a FRET-based method for MMP-9 enzyme detection. The basic aim is to apply a ratio-metric sensing technique in which a ratio of green/red fluorescence intensity is measured as a function of enzyme concentration. The ratio-metric method eliminates many experimental variables and enables accurate MMP-9 detection.

Original languageEnglish
Pages (from-to)1134-1138
Number of pages5
JournalCurrent Pharmaceutical Biotechnology
Volume14
Issue number13
DOIs
StatePublished - 1 Jan 2014

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

Keywords

  • FRET
  • Fluorescence
  • Labeled peptide
  • MMP-9
  • Ratio-metric sensing

Cite this

@article{65f9810a0703408891302595a5d26baa,
title = "Fluorescence detection of MMP-9. II. ratiometric FRET-based sensing with dually labeled specific peptide",
abstract = "In our previous paper we showed that the MMP-9 enzyme recognizes a specific peptide sequence, Lys-Gly-Pro-Arg-Ser-Leu-Ser-Gly-Lys, and cleaves the peptide into two parts [1]. In this study, the peptide is labeled with two dyes, carboxyfluorescein (5-FAM) and Cy5. A highly efficient energy transfer of over 80{\%} results in a dominant emission of Cy5 at ~670 nm with an excitation of 470 nm. Severance of the peptide by the MMP-9 enzyme eliminates F{\"o}rster Resonance Energy Transfer (FRET) and strongly increases the fluorescence of the 5-FAM dye. In this manuscript we describe the strategy for a FRET-based method for MMP-9 enzyme detection. The basic aim is to apply a ratio-metric sensing technique in which a ratio of green/red fluorescence intensity is measured as a function of enzyme concentration. The ratio-metric method eliminates many experimental variables and enables accurate MMP-9 detection.",
keywords = "FRET, Fluorescence, Labeled peptide, MMP-9, Ratio-metric sensing",
author = "Rafal Fudala and Ryan Rich and Anindita Mukerjee and Ranjan, {Amalendu Prakash} and Vishwanatha, {Jamboor K.} and Kurdowska, {Anna K.} and Zygmunt Gryczynski and Julian Borejdo and Ignacy Gryczynski",
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Fluorescence detection of MMP-9. II. ratiometric FRET-based sensing with dually labeled specific peptide. / Fudala, Rafal; Rich, Ryan; Mukerjee, Anindita; Ranjan, Amalendu Prakash; Vishwanatha, Jamboor K.; Kurdowska, Anna K.; Gryczynski, Zygmunt; Borejdo, Julian; Gryczynski, Ignacy.

In: Current Pharmaceutical Biotechnology, Vol. 14, No. 13, 01.01.2014, p. 1134-1138.

Research output: Contribution to journalArticleResearchpeer-review

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T1 - Fluorescence detection of MMP-9. II. ratiometric FRET-based sensing with dually labeled specific peptide

AU - Fudala, Rafal

AU - Rich, Ryan

AU - Mukerjee, Anindita

AU - Ranjan, Amalendu Prakash

AU - Vishwanatha, Jamboor K.

AU - Kurdowska, Anna K.

AU - Gryczynski, Zygmunt

AU - Borejdo, Julian

AU - Gryczynski, Ignacy

PY - 2014/1/1

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N2 - In our previous paper we showed that the MMP-9 enzyme recognizes a specific peptide sequence, Lys-Gly-Pro-Arg-Ser-Leu-Ser-Gly-Lys, and cleaves the peptide into two parts [1]. In this study, the peptide is labeled with two dyes, carboxyfluorescein (5-FAM) and Cy5. A highly efficient energy transfer of over 80% results in a dominant emission of Cy5 at ~670 nm with an excitation of 470 nm. Severance of the peptide by the MMP-9 enzyme eliminates Förster Resonance Energy Transfer (FRET) and strongly increases the fluorescence of the 5-FAM dye. In this manuscript we describe the strategy for a FRET-based method for MMP-9 enzyme detection. The basic aim is to apply a ratio-metric sensing technique in which a ratio of green/red fluorescence intensity is measured as a function of enzyme concentration. The ratio-metric method eliminates many experimental variables and enables accurate MMP-9 detection.

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