Increased chemical acetylation of peptides and proteins in rats after daily ingestion of diacetyl analyzed by Nano-LC-MS/MS

Abstract

BackgroundAcetylation alters several protein properties including molecular weight, stability, enzymatic activity, protein–protein interactions, and other biological functions. Our previous findings demonstrating that diacetyl/peroxynitrite can acetylate L-lysine, L-histidine, and albumin in vitro led us to investigate whether diacetyl-treated rats suffer protein acetylation as well.MethodsWistar rats were administered diacetyl daily for four weeks, after which they were sacrificed, and their lung proteins were extracted to be analysed by Nano-LC-MS/MS (Q-TOF). A C18 reversed-phase column and gradient elution with formic acid/acetonitrile solutions from 2 to 50% over 150 min were used to separate the proteins. Protein detection was performed using a microTOF-Q II (QTOF) equipped with captive source and an electrospray-ionization source. The data from mass spectrometry were processed using a Compass 1.7 and analyzed using Protein Scape, software that uses Mascot algorithms to perform protein searches.ResultsA set of 3,162 acetylated peptides derived from 351 acetylated proteins in the diacetyl-treated group was identified. Among them, 23 targeted proteins were significantly more acetylated in the diacetyl-treated group than in the PBS control. Protein acetylation of the group treated with 540 mg/kg/day of diacetyl was corroborated by Western blotting analysis.ConclusionsThese data support our hypothesis that diacetyl exposure in animals may lead to the generation of acetyl radicals, compounds that attach to proteins, affecting their functions and triggering adverse health problems.

Prevalência De Sintomas Musculoesqueléticos Em Professores Do Ensino Superior Em Uma Instituição Privada No Município De Picos-Pi

“…Assim como modificações pós traducionais causadas nas proteínas em função dessa substância. (Jedlicka et al, 2018a;Alves at al, 2013;DE CEULENEER et al, 2011) As análises proteômicas de plasma de camundongos tratados com diacetil apontaram uma diferença na expressão das proteínas entre os grupos controle e o grupo tratado com o aditivo. Na figura 1 está representada através do diagrama de Venn as alterações na expressão de proteínas entre ambos os grupos.…”

“…No organismo, o diacetil, depois de inalado, pode levar a acetilação de proteínas (Jedlicka et al, 2018a), também pode ocorrer outras modificações pós traducionais das proteínas (Alves et al, 2013), onde os radicais livres do diacetil reagem com as proteínas, alterando-as de diferentes formas, como número de massa, atividade enzimática, estabilidade, entre outras funções. Essas alterações vêm sendo elencadas como a possível causa de doenças respiratórias, como a Bronquiolite Obliterante (Cavalcanti, et al 2012).…”

“…O uso de diacetil é capaz de provocar alterações em proteínas, tanto na estrutura química como na expressão (Jedlicka et al, 2018a;Massari et al, 2011). As proteínas são as responsáveis por boa parte das atividades fisiológicas das células de um organismo, sendo de grande interesse para fins farmacológicos, uma vez que são transportadoras natas, podem contribuir expressivamente no tratamento de doenças e, como biomarcadores de doenças, podem aprimorar o método de diagnóstico, monitoramento e tratamento de patologias como o câncer, de forma precoce, ou mesmo evitar outros agravos.…”

Exceptionally versatile – arginine in bacterial post-translational protein modifications

“…Nevertheless, more research is required to unveil the physiological relevance of bacterial α-N-terminal protein acylation in general and of arginine in particular. It is worth mentioning here that there are indications coming from mass spectrometry analyses that the guanidino group of peptide bound arginine is also subject to acetylation (Jedlicka et al, 2018). However, these data so far lack biochemical proof.…”

“…In this regard, acetyltransferases catalyze the transfer of an acetyl group from an acetyl coenzyme A (acetyl CoA) donor (Christensen et al, 2019). Primarily the acyl moiety is accepted by the ε-amino group of lysine or alternatively an N-terminal α-amino group of various amino acids including arginine (Ouidir et al, 2015;Jedlicka et al, 2018). In contrast to eukaryotes, in which the majority of α-amino groups are acetylated (~80% in humans), reports of N-terminal acetylations in prokaryotes are rather scarce (Polevoda and Sherman, 2003;Jones and O'Connor, 2011;Ouidir et al, 2015).…”