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Journal Articles Proceedings Year : 2019

Design of New Probes for Oxidized Amino Acids Localization

Abstract

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. Theses probes have been explored on oxidized bovine serum albumin (OxBSA) 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.
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hal-02393325 , version 1 (05-12-2019)

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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/proceedings2019022039⟩. ⟨hal-02393325⟩
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