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 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.
authors: Center of Commercialization of Fluorescence Technologies, Department of Molecular Biology & Immunology, UNTHSC, Fort Worth, TX 76107, USA. Rafal.Fudala@unthsc.edu
authors: Rafal
authors: R
authors: Fudala
authors: Amalendu P
authors: AP
authors: Ranjan
authors: Anindita
authors: A
authors: Mukerjee
authors: Jamboor K
authors: JK
authors: Vishwanatha
authors: Zygmunt
authors: Z
authors: Gryczynski
authors: Julian
authors: J
authors: Borejdo
authors: Pabak
authors: P
authors: Sarkar
authors: Ignacy
authors: I
authors: Gryczynski
mesh_terms: D002232:Carbocyanines / Q000737:chemistry; D005453:Fluorescence; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D020778:Matrix Metalloproteinase 2 / Q000032:analysis / Q000378:metabolism; D020780:Matrix Metalloproteinase 9 / Q000032:analysis* / Q000378:metabolism; D009842:Oligopeptides / Q000378:metabolism*; D010446:Peptide Fragments / Q000032:analysis / Q000378:metabolism; D013379:Substrate Specificity
chemical_list: D002232:Carbocyanines; D009842:Oligopeptides; D010446:Peptide Fragments; C085321:cyanine dye 5; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9