Spectroscopic method for estimation of MMP-9 enzyme concentration and activity

Anna Synak , Illia Serdiuk , Beata Grobelna , Rafał Fudala , Ignacy Gryczyński , Piotr Bojarski

Abstract

Two highly sensitive fluorescent dye-labelled peptide indicators for metalloproteinase 9 (MMP-9) activity were designed and tested in vitro. Excellent sensitivity of these indicators is achieved by careful selection of the amino acid sequence corresponding to the substrate recognition preference ofMMP-9. The phenomenon of Förster resonance energy transfer (FRET) is applied for detection of the peptide cleavage by MMP-9. FRET occurs between fluorescent dyes AMCA (donor) and TAMRA (acceptor) bound to the flexible peptide fragments differing in the donor acceptor separation distance. Enzymatic hydrolysis of the peptide leads to a significant weakening of energy transfer and increased donor fluorescence intensity. The enzyme action efficiency is higher for the longer proline-rich peptide which is reflected in the results of steady-state fluorescence studies. In particular, the use of longer peptide allows shorter detection time ofMMP-9 (about 20–30 min). The possible reasons for this effect are discussed.
Author Anna Synak (FMPI / IEP)
Anna Synak,,
- Institute of Experimental Physics
, Illia Serdiuk (FMPI / IEP)
Illia Serdiuk,,
- Institute of Experimental Physics
, Beata Grobelna (FCh / DACh / LChAC)
Beata Grobelna,,
- Laboratory of chemistry and Analytics of Cosmetics
, Rafał Fudala
Rafał Fudala,,
-
, Ignacy Gryczyński
Ignacy Gryczyński,,
-
, Piotr Bojarski (FMPI / IEP)
Piotr Bojarski,,
- Institute of Experimental Physics
Journal seriesJournal of Molecular Liquids, ISSN 0167-7322, (A 30 pkt)
Issue year2019
Vol286
Pages1-5
Publication size in sheets0.5
Article number110936
Keywords in EnglishMetalloproteinase 9, FRET, fluorescence, dye-labelled peptide
ASJC Classification2505 Materials Chemistry; 1606 Physical and Theoretical Chemistry; 1607 Spectroscopy; 3104 Condensed Matter Physics; 3107 Atomic and Molecular Physics, and Optics; 2504 Electronic, Optical and Magnetic Materials
DOIDOI:10.1016/j.molliq.2019.110936
URL https://doi.org/10.1016/j.molliq.2019.110936
Languageen angielski
Score (nominal)30
ScoreMinisterial score = 30.0, 24-07-2019, ArticleFromJournal
Publication indicators Scopus SNIP (Source Normalised Impact per Paper): 2016 = 1.212; WoS Impact Factor: 2017 = 4.513 (2) - 2017=3.929 (5)
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