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Design, characterization, and evaluation of peptide arrays allowing the direct monitoring of MMP activities

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Abstract

Characterization of matrix metalloprotease (MMP) activities is of increasing interest for cancer prognosis or treatment follow-up. Indeed, MMP-1, -2 and -9 are widely involved in the growth of many tumors and progression steps such as angiogenesis, invasion, and metastasis. Fluorogenic peptide MMP substrates were previously synthesized with the aim of detecting MMP activities. One of their drawbacks is their limited solubility in biological media. Grafting them onto a solid support represented a novel way to yield efficient analysis devices whilst at the same time decreasing the quantities of peptides used. Novel peptide arrays were designed in order to detect MMP activities in biological fluids. Silicon plates were used as the solid support for the design of these novel tools. These were functionalized by organic self-assembled monolayers (SAMs) on which fluorogenic peptides were covalently coupled. SAM and peptide grafting on silicon plates were confirmed by epifluorescence, ellipsometry, and FT-IR analysis. Enzymatic assays were monitored by fluorescence spectrometry and showed that immobilized linear peptides were recognized and cleaved by MMPs.

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Abbreviations

DAC:

7-diethylamino coumarin-3-carboxylic acid

DCM:

methylene chloride

DIC:

N,N′-diisopropyl carbodiimide

DIEA:

N,N′-diisopropydiethylamine

DMF:

N,N-dimethylformamide

ECM:

extracellular matrix

FRET:

fluorescence resonance energy transfer

FT-IR:

Fourier-transform infrared

HOBt:

N-hydroxybenzotriazole

MC:

7-methoxy coumarin-3-carboxylic acid

MMP:

matrix metalloprotease

MS:

mass spectrometry

SAM:

self assembled monolayer

TEG:

triethyleneglycol

THF:

tetrahydrofuran

TLC:

thin layer chromatography

TRIS:

tris(hydroxymethyl)aminomethane

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Acknowledgments

The authors wish to warmly thank Dr. Bernard Desbat (CNRS Bordeaux) for helpful discussions and technical support for ellipsometry analysis and the Conseil Régional d’Aquitaine and Ligue Nationale Contre le Cancer for financial support. Careful reading and corrections of our manuscript by Dr. Robert Dodd are greatly acknowledged.

Author information

Authors and Affiliations

  1. Université Bordeaux Segalen, CNRS, FRE 3396 Pharmacochimie, 146 rue Léo Saignat, 33076, Bordeaux Cedex, France

    Mohamed-Anis Alouini, El-Farouck Moustoifa, Sandra Albenque Rubio & Gérard Déléris

  2. Chemistry of Surfaces and Interfaces CEA Saclay, Bâtiment 466, F-91191, Gif-sur-Yvette, CEDEX, France

    Thomas Berthelot

  3. Unité de Recherche 02/UR/09-01 Biochimie des Protéines et Interactions Moléculaires, Avenue Taher Hadda, BP 74, Monastir, 5000, Tunisia

    Mohamed-Anis Alouini & Aghleb Bartegi

Authors
  1. Mohamed-Anis Alouini
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  2. El-Farouck Moustoifa
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  3. Sandra Albenque Rubio
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  4. Aghleb Bartegi
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  5. Thomas Berthelot
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  6. Gérard Déléris
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Correspondence to Mohamed-Anis Alouini.

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Professor Déléris regrettably deceased in January 2012.

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Alouini, MA., Moustoifa, EF., Rubio, S.A. et al. Design, characterization, and evaluation of peptide arrays allowing the direct monitoring of MMP activities. Anal Bioanal Chem 403, 185–194 (2012). https://doi.org/10.1007/s00216-012-5760-x

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  • DOI: https://doi.org/10.1007/s00216-012-5760-x

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