For these reasons, protein carbonyl content is widely used as a marker of protein oxidative damage, including for age-related disorders. In comparison, carbonyl modification is irreversible and therefore causes permanent damage to proteins. However, these oxidative modifications can be repaired by the cell and most biological systems contain enzymes (disulfide reductases and MeSOX reductases) that can convert the oxidized forms of cysteine and methionine residues, respectively, back to their unmodified forms. Sulfur-containing amino acids such as cysteine and methionine are also susceptible to ROS. Oxidative stress often leads to proteins becoming modified by carbonylation. Carbonyl groups cause changes in protein hydrophobicity, surface charge and associated misfolding of proteins. Oxidative stress arises when reactive oxygen species (ROS) accumulate in cells to the extent that DNA, RNA, lipids and proteins are damaged by oxidation.Ĭarbonylation is the post-translational addition of a carbonyl group (C = O) to the side chains of amino acids and preferentially affects arginine, threonine, proline or lysine ( Figure 1), and is caused by diverse oxidative reactions, in particular by direct metal-catalyzed oxidative attack. Damage includes the dimerization of two adjacent pyrimidine bases located on the same strand of DNA and/or indirect oxidative stress. The increased UVB irradiation constitutes a potentially important risk to many life forms as this type of radiation damages many different cellular components. The discovery of the ‘ozone hole’ over Antarctica in the 1980’s prompted research into the environmental impacts of increasing levels of solar ultraviolet radiation, particularly UVB (280–320 nm), reaching the Earth’s surface. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.Ĭompeting interests: The authors have declared that no competing interests exist. Research in RC and MJRs laboratories was supported by the Australian Research Council. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.įunding: This work was supported by the European community project MAMBA (FP7-KBBE-2008-226977). Received: ApAccepted: Published: July 9, 2013Ĭopyright: © 2013 Matallana-Surget et al. PLoS ONE 8(7):Įditor: Maria Gasset, Consejo Superior de Investigaciones Cientificas, Spain (2013) Shotgun Redox Proteomics: Identification and Quantitation of Carbonylated Proteins in the UVB-Resistant Marine Bacterium, Photobacterium angustum S14.
As the first large scale, shotgun redox proteomics analysis examining carbonylation to be performed on bacteria, our study provides a new level of understanding about the effects of UVB on cellular proteins, and provides a methodology for advancing studies in other biological systems.Ĭitation: Matallana-Surget S, Cavicchioli R, Fauconnier C, Wattiez R, Leroy B, Joux F, et al. As a result we determined which functional classes of proteins were carbonylated, which residues were preferentially modified, and what the implications of the carbonylation were for protein function. Eleven carbonylated proteins were quantified and the UVB/dark abundance ratio was determined at both the protein and peptide levels. The DNPH redox proteomics method enabled the identification of 62 carbonylated proteins (5% of 1221 identified proteins) in cells exposed to UVB or darkness. Mass-spectrometry was performed with either biotin-labeled or dinitrophenylhydrazide (DNPH) derivatized proteins. In this study, redox proteomics was performed to identify carbonylated proteins in the UVB resistant marine bacterium Photobacterium angustum. One consequence of UVB irradiation is carbonylation, the irreversible formation of a carbonyl group on proline, lysine, arginine or threonine residues. UVB oxidizes proteins through the generation of reactive oxygen species.
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