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.
Keywords: Fluorescence, FRET, labeled peptide, MMP-9, ratio-metric sensing.
Current Pharmaceutical Biotechnology
Title:Fluorescence Detection of MMP-9. II. Ratiometric FRET-Based Sensing With Dually Labeled Specific Peptide
Volume: 14 Issue: 13
Author(s): Rafal Fudala, Ryan Rich, Anindita Mukerjee, Amalendu P. Ranjan, Jamboor K. Vishwanatha, Anna K. Kurdowska, Zygmunt Gryczynski, Julian Borejdo and Ignacy Gryczynski
Affiliation:
Keywords: Fluorescence, FRET, labeled peptide, MMP-9, ratio-metric sensing.
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.
Export Options
About this article
Cite this article as:
Fudala Rafal, Rich Ryan, Mukerjee Anindita, Ranjan P. Amalendu, Vishwanatha K. Jamboor, Kurdowska K. Anna, Gryczynski Zygmunt, Borejdo Julian and Gryczynski Ignacy, Fluorescence Detection of MMP-9. II. Ratiometric FRET-Based Sensing With Dually Labeled Specific Peptide, Current Pharmaceutical Biotechnology 2013; 14 (13) . https://dx.doi.org/10.2174/138920101413140605111109
DOI https://dx.doi.org/10.2174/138920101413140605111109 |
Print ISSN 1389-2010 |
Publisher Name Bentham Science Publisher |
Online ISSN 1873-4316 |
- Author Guidelines
- Graphical Abstracts
- Fabricating and Stating False Information
- Research Misconduct
- Post Publication Discussions and Corrections
- Publishing Ethics and Rectitude
- Increase Visibility of Your Article
- Archiving Policies
- Peer Review Workflow
- Order Your Article Before Print
- Promote Your Article
- Manuscript Transfer Facility
- Editorial Policies
- Allegations from Whistleblowers
Related Articles
-
Neoadjuvant Chemotherapy Before Radical Cystectomy: Why We Must Adhere?
Current Drug Targets New Potential Pharmaceutical Applications of Hypericum Species
Mini-Reviews in Medicinal Chemistry Heat Shock Protein 70s as Potential Molecular Targets for Colon Cancer Therapeutics
Current Medicinal Chemistry Angiotensin II Receptor Blocker: Possibility of Antitumor Agent for Prostate Cancer
Mini-Reviews in Medicinal Chemistry Withdrawal Notice: The Recent Advancement in the Field of Super Paramagnetic Iron Oxide Nanoparticles (SPIONs) for Aiming Breast Cancer
Current Drug Metabolism Epigenetics: Relations to Disease and Laboratory Findings
Current Medicinal Chemistry Meitner-Auger Electron Emitters for Targeted Radionuclide Therapy: Mercury-197m/g and Antimony-119
Current Radiopharmaceuticals The Wnt/Beta-Catenin Pathway in Wilms Tumors and Prostate Cancers
Current Molecular Medicine Biomarkers for VEGF-Targeted Therapy in Renal Cell Carcinoma: Critical Review of Level of Evidence and Up to Date
Recent Patents on Biomarkers Tumor Specific Imaging Using Tc-99m and Ga-68 Labeled Radiopharmaceuticals
Current Medical Imaging The Role of Protein Kinase C-alpha (PKC-α) in Cancer and its Modulation by the Novel PKC-α-Specific Inhibitor Aprinocarsen
Current Pharmaceutical Design Purinergic (P2) Receptor Control of Lower Genitourinary Tract Function and New Avenues for Drug Action: An Overview
Current Pharmaceutical Design Current Perspectives on Novel Drug Delivery Systems and Therapies for Management of Prostate Cancer: An Inclusive Review
Current Drug Targets The Role of CD40 Expression in Dendritic Cells in Cancer Biology; A Systematic Review
Current Cancer Drug Targets Botulinum Toxin a in Prostate Disease: A Venom from Bench to Bed-Side
Current Drug Delivery A Potencial Theranostic Agent for EGF-R Expression Tumors: 177Lu- DOTA-Nimotuzumab
Current Radiopharmaceuticals Intravesical Bacillus Calmette-Guerin Therapy for Bladder Cancer: Molecular Mechanisms of Action
Clinical Immunology, Endocrine & Metabolic Drugs (Discontinued) Management of Inflammatory Bowel Disease Patients with a Cancer History
Current Drug Targets Marine Metabolites Overcoming or Circumventing Multidrug Resistance Mediated by ATP-Dependent Transporters: A New Hope for Patient with Tumors Resistant to Conventional Chemotherapy
Anti-Cancer Agents in Medicinal Chemistry DNA Methyltransferases Inhibitors from Natural Sources
Current Topics in Medicinal Chemistry