FRET study in oligopeptide-linked donor-acceptor system in PVA matrix

Sunil Shah, Wlodek Mandecki, Ji Li, Zygmunt Gryczynski, Julian Borejdo, Ignacy Gryczynski, Rafal Fudala

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An oligopeptide: Lys-Gly-Pro-Arg-Ser-Leu-Ser-Gly-Lys-NH2, cleaved specifically by a matrix metalloproteinase 9 (MMP-9) at the Ser-Leu bond, was labeled on the ε-NH2 groups of lysine with donor (5, 6 TAMRA) and acceptor (HiLyte647) dye. The donor control was a peptide labeled with 5, 6 TAMRA only on the C-terminal lysine, and the acceptor control was free HiLyte647. Following three products were studied by dissolving in 10% (w/w) poly(vinyl alcohol) and dried on glass slides forming 200 micron films. Absorption spectra of the films show full additivity of donor and acceptor absorptions. A strong Fluorescence Resonance Energy Transfer (FRET) with an efficiency of about 85% was observed in the fluorescence emission and excitation spectra. The lifetime of the donor was shorter and heterogeneous compared with the donor control.

Original languageEnglish
Number of pages1
JournalMethods and Applications in Fluorescence
Issue number4
StatePublished - 13 Dec 2016


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