Shotgun Redox Proteomics: Identification and Quantitation of Carbonylated Proteins in the UVB-Resistant Marine Bacterium, Photobacterium angustum S14

PLoS One

Baptiste Leroy¹  Ricardo Cavicchioli¹  Fabien Joux²  Mark J. Raftery²  Charles Fauconnier³  Ruddy Wattiez⁴  Sabine Matallana-Surget⁴  Philippe Lebaron⁴ 

[1] CNRS, UMR7621, Laboratoire d’Océanographie Microbienne, Observatoire Océanologique, Banyuls/mer, France;Department of Proteomics and Microbiology, Research Institute for Biosciences Interdisciplinary Mass Spectrometry Center (CISMa), University of Mons, Mons, Belgium;School of Biotechnology and Biomolecular Sciences, The University of New South Wales, Sydney, Australia;UPMC Univ Paris 06, UMR7621, Laboratoire d’Océanographie Microbienne, Observatoire Océanologique, Banyuls/mer, France

关键词: Chaperone proteins;    Biotin;    Ultraviolet B;    Proteomics;    Elongation factors;    Oxidation-reduction reactions;    Marine bacteria;    Arginine;   

DOI  :  10.1371/journal.pone.0068112

学科分类：医学（综合）

来源: Public Library of Science

【 摘 要 】

UVB oxidizes proteins through the generation of reactive oxygen species. One consequence of UVB irradiation is carbonylation, the irreversible formation of a carbonyl group on proline, lysine, arginine or threonine residues. In this study, redox proteomics was performed to identify carbonylated proteins in the UVB resistant marine bacterium Photobacterium angustum. Mass-spectrometry was performed with either biotin-labeled or dinitrophenylhydrazide (DNPH) derivatized proteins. The DNPH redox proteomics method enabled the identification of 62 carbonylated proteins (5% of 1221 identified proteins) in cells exposed to UVB or darkness. Eleven carbonylated proteins were quantified and the UVB/dark abundance ratio was determined at both the protein and peptide levels. 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. 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.

【 授权许可】

CC BY