# Design of New Probes for Oxidized Amino Acids Localization

Mathieu Esgulian, Luc Camoin, Mathieu Cassien, Yves Toiron, Sylvia Pietri,

Sophie Thétiot-Laurent

► To cite this version:

Mathieu Esgulian, Luc Camoin, Mathieu Cassien, Yves Toiron, Sylvia Pietri, et al. Design of New Probes for Oxidized Amino Acids Localization. Proceedings, 2019, 22 (1), pp.39. 10.3390/proceed-ings2019022039. [https://arxiv.org/abs/1902.03257](https://arxiv.org/abs/1902.03257)

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Abstract

# Design of New Probes for Oxidized Amino Acids Localization †

Mathieu Esgulian 1,*, Luc Camoin 2, Mathieu Cassien 1, Yves Toiron 2, Sylvia Pietri 1 and
Sophie Thétiot-Laurent 1,*

Published: 8 August 2019

**Keywords:** protein carbonyls; probes; hydrazine; click chemistry

Protein carbonyls (PC) are oxidative damage observed in many diseases. Proteins are possibly the most immediate vehicle for inflicting oxidative damage on cells because they are often catalysts [1]. A fluorometric and UV-absorption method have been developed to quantify PC in blood and tissues samples by labeling with two hydrazines: 7-hydrazino-4-nitrobenzo-2,1,3-oxadiazole (NBDH) and dinitrophenylhydrazine (DNPH), respectively [2,3]. These methods are based on the selective hydrazone formation between the carbonyls group of oxidized protein to yield a strong fluorescent/UV-absorption adduct that is then quantified.

Here, we will describe our study on NBDH's and DNPH's derivatives to generate new PC-probes bearing an alkyne moiety. In a first step, the hydrazine moiety reacted specifically with protein carbonyls and in a second step, a click reaction was performed between the alkyne moiety and a cleavable resin [4] to yield a PC's enrichment. These probes have been explored on oxidized bovine serum albumin (OBSA) for PC-labeling, and the possible sites of oxidation of isolated labelled PC will be studied by LC-MS and proteomics experiments.

## References

1. Dalle-Donne, I.; Rossi, R.; Giustarini, D.; Milzani, A.; Colombo, R. Protein carbonyl groups as biomarkers of oxidative stress. *Clin. Chim. Acta* **2003**, *329*, 23–38.

2. Stocker, P.; Ricquebourg, E.; Vidal, N.; Villard, C.; Lafitte, D.; Sellami, L.; Pietri, S. Fluorimetric screening assay for protein carbonyl evaluation in biological samples. *Anal. Biochem.* **2015**, *482*, 55–61.

3. Vidal, N.; Cavaille, J.P.; Graziani, F.; Robin, M.; Ouari, O.; Stocker, P. High throughput assay for evaluation of reactive carbonyl scavenging capacity. *Redox Biology* **2014**, *2*, 590–598.

4. Sibbersen, C.; Lykke, L.; Gregersen, N.; Jørgensen, K.A.; Johannsen, M. A cleavable azide resin for direct click chemistry mediated enrichment of alkyne-labeled proteins. *Chem. Commun.* **2014**, *50*, 12098–12100.

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© 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

<footnote n="1">¹ Aix Marseille Université, CNRS UMR 7273, Institut de Chimie Radicalaire, Equipe Sonde Moléculaires en Biologie et Stress Oxydant, 13013 Marseille, France</footnote>

<footnote n="2">² CRCM, Inserm, 13009 Marseille, France</footnote>

<footnote>* Correspondence: mathieu.esgulian@univ-amu.fr (M.E.); sophie.thetiot-laurent@univ-amu.fr (S.T.-L.)</footnote>

<footnote>† Presented at the 2nd Molecules Medicinal Chemistry Symposium (MMCS): Facing Novel Challenges in Drug Discovery, Barcelona, Spain, 15–17 May 2019.</footnote>