期刊
JOURNAL OF PROTEOMICS
卷 74, 期 12, 页码 2798-2809出版社
ELSEVIER
DOI: 10.1016/j.jprot.2011.06.014
关键词
Ostreococcus tauri; Unicellular algae; Glufosinate; Herbicide effect; Acetaldehyde; Reductive amine
资金
- Centre for Systems Biology at Edinburgh (CSBE), a Centre for Integrative Systems Biology (CISB)
- BBSRC
- EPSRC [BB/D019621/1]
- Biotechnology and Biological Sciences Research Council [BB/D019621/1] Funding Source: researchfish
- BBSRC [BB/D019621/1] Funding Source: UKRI
We report a novel stable-isotope labeling strategy for quantitative proteomics analysis. The method consists of labeling N-termini and lysine c-amino groups through reductive amination using acetaldehyde. This allows isotope labeling using pairs of either H-2/H-1 or C-13/C-12 without mass spectrum overlap. Our labeling procedure, which is significantly different than that developed for dimethylation, can be completed with little trace of partial ethylation; non-labeled peptides represent less than 0.05% of all peptides. Co-elution of both isotopic C-13/C-12 peptide pairs was observed in all cases, simplifying data analysis, which can be performed using standard commercial software such as Mascot Distiller. A C-13/C-12 labeled mix in a 1:1 ratio from a complex extract digest of the unicellular algae Ostreococcus tauri, showed a relative standard deviation of less than 14%. This quantitative method was used to characterize O. tauri in the presence of glufosinate, an herbicide which inhibits glutamine synthetase. Blocking glutamine synthetase significantly reduced the expression of several enzymes and transporters involved in nitrogen assimilation and the expression of a number of proteins involved in various stresses including oxidative damage response were up-regulated. (C) 2011 Elsevier B.V. All rights reserved.
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