Increased chemical acetylation of peptides and proteins in rats after daily ingestion of diacetyl analyzed by Nano-Lc-MS/MS
- Published
- Accepted
- Subject Areas
- Biochemistry, Toxicology
- Keywords
- Radical acetylation, diacetyl, food additive, lung diseases, proteomics.
- Copyright
- © 2018 Jedlicka et al.
- Licence
- This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, reproduction and adaptation in any medium and for any purpose provided that it is properly attributed. For attribution, the original author(s), title, publication source (PeerJ Preprints) and either DOI or URL of the article must be cited.
- Cite this article
- 2018. Increased chemical acetylation of peptides and proteins in rats after daily ingestion of diacetyl analyzed by Nano-Lc-MS/MS. PeerJ Preprints 6:e26613v1 https://doi.org/10.7287/peerj.preprints.26613v1
Abstract
Background: Acetylation 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.
Methods: Wistar rats were administered diacetyl daily for 4 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.
Results: A set of 3162 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.
Conclusions: These 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.
Author Comment
This is a submission to PeerJ for review.
Supplemental Information
NanoHPLC gradient elution
NanoHPLC gradient elution, gradient mobile phase consisting of (A) formic acid/ACN/ H2O (1:20:980) and (B) formic acid/ACN/ H2O (1:950:50)
Central residue H acetylated (control group)
Motif Analyse of Lung from control group.Central residue H acetylated
Central residue H acetylated in group treated with 540 mg/Kg/day of Diacetyl
Motif Analyse of Lung from group treated with 540 mg/Kg/day of Diacetyl. Central residue H acetylated
Central residue K acetylated in group treated with 540 mg/Kg/day of Diacetyl
Motif Analyse of Lung from group treated with 540 mg/Kg/day of Diacetyl. Central residue K acetylated
Central residue K acetylated in control group
Motif Analyse of Lung from control group. Central residue K acetylated
Central residue R acetylated in group treated with 540 mg/Kg/day of Diacetyl
Mottif Analyse of Lung from group treated with 540 mg/Kg/day of Diacetyl. Central residue R acetylated
Central residue R acetylated in control group
Motif Analyse of Lung from control group. Central residue R acetylated