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 |
Call for Papers in Thematic Issues
Artificial Intelligence in Bioinformatics
Bioinformatics is an interdisciplinary field that analyzes and explores biological data. This field combines biology and information system. Artificial Intelligence (AI) has attracted great attention as it tries to replicate human intelligence. It has become common technology for analyzing and solving complex data and problems and encompasses sub-fields of machine ...read more
- 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
-
Folate Based Radiopharmaceuticals for Imaging and Therapy of Cancer and Inflammation
Current Pharmaceutical Design Intrathecal Administration of Melatonin Ameliorates the Neuroinflammation- Mediated Sensory and Motor Dysfunction in A Rat Model of Compression Spinal Cord Injury
Current Molecular Pharmacology Targeting Metallothionein for Prognosis and Treatment of Breast Cancer
Recent Patents on Anti-Cancer Drug Discovery The Role of Mass Spectrometry in the “Omics” Era
Current Organic Chemistry The Role of TRP Channels in Allergic Inflammation and its Clinical Relevance
Current Medicinal Chemistry Antidiabetic Potential of Fabaceae Family: An Overview
Current Nutrition & Food Science Anticancer α-Helical Peptides and Structure / Function Relationships Underpinning Their Interactions with Tumour Cell Membranes
Current Protein & Peptide Science New Approaches With Natural Product Drugs for Overcoming Multidrug Resistance in Cancer
Current Pharmaceutical Design Signaling Pathways Underpinning the Manifestations of Ionizing Radiation-Induced Bystander Effects
Current Molecular Pharmacology Progresses in TCM Metal-Based Antitumour Agents
Anti-Cancer Agents in Medicinal Chemistry Doxorubicin-Induced In Vivo Nephrotoxicity Involves Oxidative Stress- Mediated Multiple Pro- and Anti-Apoptotic Signaling Pathways
Current Neurovascular Research Curcumin Targets Circulating Cancer Stem Cells by Inhibiting Self-Renewal Efficacy in Non-Small Cell Lung Carcinoma
Anti-Cancer Agents in Medicinal Chemistry Lymphoblastoid Cell Lines Models of Drug Response: Successes and Lessons from this Pharmacogenomic Model
Current Molecular Medicine Targeting Glycosylation Aberrations to Improve the Efficiency of Cancer Phototherapy
Current Cancer Drug Targets Role of Neuromodulation and Optogenetic Manipulation in Pain Treatment
Current Neuropharmacology Radioactive Gold Nanoparticle in Two Forms (<sup>198</sup><sub>79</sub>Au GNPs and <sup>99m</sup>Tc-GNPs) for Lung Cancer Antiproliferative Induction and Intralesional Imaging: A Proof of Concept
Anti-Cancer Agents in Medicinal Chemistry Assortment of Human Amniotic Membrane and Curcumin: a Potential Therapeutic Strategy for Burn Wound Healing
Current Drug Therapy Cerebral Angiogenesis and Expression of Angiogenic Factors in Aging Rats after Exercise
Current Neurovascular Research Prognostic Value of MiRNAs in Patients with Laryngeal Cancer: A Systematic Review and Meta-Analysis
Current Cancer Drug Targets Current and Future Clinical Strategies in Colon Cancer Prevention and the Emerging Role of Chemoprevention
Current Pharmaceutical Design