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

A. Synak, I. E. Serdiuk, B. Grobelna, Rafal Fudala, Ignacy Gryczynski, P. Bojarski

Research output: Contribution to journalArticleResearchpeer-review

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 of MMP-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 of MMP-9 (about 20–30 min). The possible reasons for this effect are discussed.

Original languageEnglish
Article number110936
JournalJournal of Molecular Liquids
Volume286
DOIs
StatePublished - 15 Jul 2019

Fingerprint

Matrix Metalloproteinases
Peptides
peptides
enzymes
Enzymes
Energy transfer
Fluorescent Dyes
energy transfer
Fluorescence
Tranexamic Acid
Dyes
dyes
Peptide Fragments
Enzymatic hydrolysis
Matrix Metalloproteinase 9
fluorescence
Proline
amino acids
hydrolysis
Amino acids

Keywords

  • Dye-labelled peptide
  • FRET
  • Fluorescence
  • Metalloproteinase 9

Cite this

@article{c5cf63b43f3a4f309357290604f8d9d6,
title = "Spectroscopic method for estimation of MMP-9 enzyme concentration and activity",
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 of MMP-9. The phenomenon of F{\"o}rster 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 of MMP-9 (about 20–30 min). The possible reasons for this effect are discussed.",
keywords = "Dye-labelled peptide, FRET, Fluorescence, Metalloproteinase 9",
author = "A. Synak and Serdiuk, {I. E.} and B. Grobelna and Rafal Fudala and Ignacy Gryczynski and P. Bojarski",
year = "2019",
month = "7",
day = "15",
doi = "10.1016/j.molliq.2019.110936",
language = "English",
volume = "286",
journal = "Journal of Molecular Liquids",
issn = "0167-7322",
publisher = "Elsevier",

}

Spectroscopic method for estimation of MMP-9 enzyme concentration and activity. / Synak, A.; Serdiuk, I. E.; Grobelna, B.; Fudala, Rafal; Gryczynski, Ignacy; Bojarski, P.

In: Journal of Molecular Liquids, Vol. 286, 110936, 15.07.2019.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - Spectroscopic method for estimation of MMP-9 enzyme concentration and activity

AU - Synak, A.

AU - Serdiuk, I. E.

AU - Grobelna, B.

AU - Fudala, Rafal

AU - Gryczynski, Ignacy

AU - Bojarski, P.

PY - 2019/7/15

Y1 - 2019/7/15

N2 - 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 of MMP-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 of MMP-9 (about 20–30 min). The possible reasons for this effect are discussed.

AB - 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 of MMP-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 of MMP-9 (about 20–30 min). The possible reasons for this effect are discussed.

KW - Dye-labelled peptide

KW - FRET

KW - Fluorescence

KW - Metalloproteinase 9

UR - http://www.scopus.com/inward/record.url?scp=85065544923&partnerID=8YFLogxK

U2 - 10.1016/j.molliq.2019.110936

DO - 10.1016/j.molliq.2019.110936

M3 - Article

VL - 286

JO - Journal of Molecular Liquids

JF - Journal of Molecular Liquids

SN - 0167-7322

M1 - 110936

ER -