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Use of proteomics techniques to study radical acetylation of proteins triggered by products of the reaction biacetyl/peroxinitrite

Abstract

This project aims to use bioanalytical tools in order to study the radical acetylation of the proteins. The superoxide anion is produced in oxidative stress and reacts with nitric oxide generating peroxynitrite which attacks the alpha carbon of the diacetyl producing radical species. These radicals are able to acetylate proteins and nitrogenous bases in vitro. The exposure to diacetyl is associated to several illnesses, mostly lung diseases. The non controled radical acetylation may cause changes in protein functions and genetic failure which implies in diseases and mutation. The purpose of this work is to verify the radical acetylation caused by peroxinitrite/diacetyl reaction in vivo and the influence of these changes in the proteomic expression in lung tissue. It will be carried out a study using Wistar mice treated with i) inactive ingredient, ii) peroxynitrite; iii) 13C labeled diacetyl; iv) 13C labeled diacetyl and peroxynitrite. The metabolites and proteins extracted from each group will be analyzed by capillary electrophoresis, two dimensional gel electrophoresis, liquid chromatography, western blot and mass spectrometry in order to separate and identify proteins and the position of their acetylation. From these results we hope to answer some relevant biological questions proposing a non enzymatic mechanism to protein acetylation and how it is related with the proteomic expression in oxidative stress and the exposure of the diacetyl. The answers to these questions may contribute to understand the origin of the several illnesses and it will drive the need to control the use of diacetyl in food. (AU)

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Scientific publications (10)
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
TRINTINALIA, MAIRA MAGALHAES; LOPES ALVES, ATECLA NUNCIATA; FERNANDES, LILIAM; HENRIQUES BECHARA, ETELVINO JOSE; ASSUNCAO, NILSON ANTONIO. Potential diagnostic of Branched-Chain Ketoaciduria by HPLC-DAD. Journal of the Brazilian Chemical Society, v. 25, n. 10, p. 1857-1863, . (12/02514-9)
FIGUEIREDO LEITE, GIUSEPPE GIANINI; AZEVEDO, HATYLAS; DE OLIVEIRA, TALITA MENDES; SOUSA FURTADO, DANIELLE ZILDEANA; ASSUNCAO, NILSON ANTONIO. Cri-du-Chat Syndrome interactome network: Correlating genotypic variations to associated phenotypes. GENE REPORTS, v. 11, p. 179-187, . (13/21455-6, 12/02514-9)
LIMA JEDLICKA, LETICIA DIAS; GUTERRES, SHEILA BARRETO; BALBINO, ALEKSANDRO MARTINS; NETO, GIUSEPPE BRUNO; LANDGRAF, RICHARDT GAMA; FERNANDES, LILIAM; CARRILHO, EMANUEL; HENRIQUES BECHARA, ETELVINO JOSE; ASSUNCAO, NILSON A.. Increased chemical acetylation of peptides and proteins in rats after daily ingestion of diacetyl analyzed by Nano-LC-MS/MS. PeerJ, v. 6, . (13/07763-0, 12/02514-9, 10/01404-0)
SILVA, ADRIANA R.; DE LACERDA, JOSE T. J. G.; FARIA, BERNADETE; DA CUNHA, ISABELA W.; DE ANDRADE, VITOR P.; ASSUNCAO, NILSON A.. Morpho/Proteomic Comparative between High Grade Pleomorphic Sarcoma and Metastasis Diagnosed in an Old Captive Common Hippo. Journal of the Brazilian Chemical Society, v. 29, n. 2, p. 363-370, . (12/02514-9)
JOCELINO GOMES DE LACERDA, JOSE THALLES; RODRIGUES E LACERDA, RODRIGO; ASSUNCAO, NILSON ANTONIO; TASHIMA, ALEXANDRE KEIJI; JULIANO, MARIA APARECIDA; DOS SANTOS, JR., GILBERTO ALVES; DE SOUZA, MILENY DOS SANTOS; BATISTA, JACINTO DE LUNA; ROSSI, CARLOS EDUARDO; DE ALMEIDA GADELHA, CARLOS ALBERTO; et al. New insights into lectin from Abelmoschus esculentus seeds as a Kunitz-type inhibitor and its toxic effects on Ceratitis capitata and root-knot nematodes Meloidogyne spp.. Process Biochemistry, v. 63, p. 96-104, . (12/19321-9, 12/02514-9, 12/50191-4)
LIMA JEDLICKA, LETICIA DIAS; SILVA, JUCIARA DA COSTA; BALBINO, ALEKSANDRO MARTINS; NETO, GIUSEPPE BRUNO; SOUSA FURTADO, DANIELLE ZILDEANA; TORRES DA SILVA, HERON DOMINGUEZ; CORREIA CAVALCANTI, FERNANDADE BARROS; VAN DER HEIJDEN, KARIN MARIE; AVELLANEDA PENATTI, CARLOS ALBERTO; HENRIQUES BECHARA, ETELVINO JOSE; et al. Effects of Diacetyl Flavoring Exposure in Mice Metabolism. BIOMED RESEARCH INTERNATIONAL, . (12/02514-9, 18/02385-0, 13/07763-0)
SOUSA FURTADO, DANIELLE ZILDEANA; VILELA DE MOURA LEITE, FERNANDO BRUNALE; BARRETO, CLEBER NUNES; FARIA, BERNADETE; LIMA JEDLICKA, LETICIA DIAS; SILVA, ELISANGELA DE JESUS; TORRES DA SILVA, HERON DOMINGUEZ; HENRIQUES BECHARA, ETELVINO JOSE; ASSUNCAO, NILSON ANTONIO. Profiles of amino acids and biogenic amines in the plasma of Cri-du-Chat patients. Journal of Pharmaceutical and Biomedical Analysis, v. 140, p. 137-145, . (13/07763-0, 13/21455-6, 12/02514-9)
FURTADO, DANIELLE ZILDEANA SOUSA; LEITE, FERNANDO BRUNALE VILELA DE MOURA; JEDLICKA, LETICIA DIAS LIMA; SOUZA, DANILO SANTOS; BARRETO, CLEBER NUNES; DA SILVA, HERON DOMINGUEZ TORRES; ASSUNCAO, NILSON ANTONIO. Targeted analysis reveals alteration in pathway in 5p minus individuals. BIOMEDICAL CHROMATOGRAPHY, v. 34, n. 1, . (18/02385-0, 12/02514-9, 13/21455-6)
LIMA JEDLICKA, LETICIA DIAS; SILVA, JUCIARA DA COSTA; BALBINO, ALEKSANDRO MARTINS; NETO, GIUSEPPE BRUNO; SOUSA FURTADO, DANIELLE ZILDEANA; TORRES DA SILVA, HERON DOMINGUEZ; CORREIA CAVALCANTI, FERNANDADE BARROS; VAN DER HEIJDEN, KARIN MARIE; AVELLANEDA PENATTI, CARLOS ALBERTO; HENRIQUES BECHARA, ETELVINO JOSE; et al. Effects of Diacetyl Flavoring Exposure in Mice Metabolism. BIOMED RESEARCH INTERNATIONAL, v. 2018, p. 11-pg., . (13/07763-0, 18/02385-0, 12/02514-9)
ALVES, ATECLA N. L.; JEDLICKA, LETICIA D. L.; MASSARI, JULIO; JULIANO, MARIA A.; BECHARA, ETELVINO J. H.; ASSUNCAO, NILSON A.. Electrospray ionization mass spectrometry applied to study the radical acetylation of amino acids, peptides and proteins. Journal of the Brazilian Chemical Society, v. 24, n. 12, p. 1983+, . (12/02514-9)

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