Spectroscopic method for estimation of MMP-9 enzyme concentration and activity
Anna Synak , Illia Serdiuk , Beata Grobelna , Rafał Fudala , Ignacy Gryczyński , Piotr Bojarski
AbstractTwo 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.
|Journal series||Journal of Molecular Liquids, ISSN 0167-7322, (A 30 pkt)|
|Publication size in sheets||0.5|
|Keywords in English||Metalloproteinase 9, FRET, fluorescence, dye-labelled peptide|
|ASJC Classification||; ; ; ; ;|
|Score||= 30.0, 24-07-2019, ArticleFromJournal|
|Publication indicators||: 2016 = 1.212; : 2017 = 4.513 (2) - 2017=3.929 (5)|
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