Abstract
Leptospirosis is a widespread zoonotic disease caused by pathogenic spirochetes of the genus Leptospira that infects humans and a wide range of animals. By combining computational prediction and high-accuracy tandem mass spectra, we revised the genome annotation of Leptospira interrogans serovar Lai, a free-living pathogenic spirochete responsible for leptospirosis, providing substantial peptide evidence for novel genes and new gene boundaries. Subsequently, we presented a high-coverage proteome analysis of protein expression and multiple posttranslational modifications (PTMs). Approximately 64.3% of the predicted L. interrogans proteins were cataloged by detecting 2 540 proteins. Meanwhile, a profile of multiple PTMs was concurrently established, containing in total 32 phosphorylated, 46 acetylated and 155 methylated proteins. The PTM systems in the serovar Lai show unique features. Unique eukaryotic-like features of L. interrogans protein modifications were demonstrated in both phosphorylation and arginine methylation. This systematic analysis provides not only comprehensive information of high-coverage protein expression and multiple modifications in prokaryotes but also a view suggesting that the evolutionarily primitive L. interrogans shares significant similarities in protein modification systems with eukaryotes.
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Acknowledgements
This work was supported by the National Natural Science Foundation of China (30425021, 30521005, 30670102, 30770111, 30770820), the Basic Research Foundation (2006CB910700), the CAS Project (KSCX2-YW-R-106, KSCX1-YW-02), the High-technology Project (2007AA02Z334) and the National High Technology Research and Development Program of China (2006AA02Z176).
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Supplementary information
Supplementary information, Table S1
Ten MS-detected proteins that were missed in computational prediction. The chromosomal location includes the stop codon. (XLS 21 kb)
Supplementary information, Table S2
MS-detected proteins of L. interrogans strain Lai in our study. (XLS 564 kb)
Supplementary information, Table S4
Evolutionary conservation of 3 953 predicted proteins of L. interrogans strain Lai. The 49 bacterial, ten archaeal and ninw eukaryotic species listed in the row 1 are used to perform the evolutionary analysis. “2”, “1” and “0” denote two-dimensional homolog, one-dimensional homolog and no homolog respectively. (XLS 6556 kb)
Supplementary information, Figure S1A, Figure S1B, Figure S1C, Figure S1D
Phosphopeptide spectra (Page 2–35), Acetylpeptide spectra(Page 36–82), Glx-methylated peptide spectra(Page 83–203), K/R-methylated peptide spectra (Page 204–273) (PDF 53508 kb)
Supplementary information, Figure S2
Pro-Q staining and Coomassie Brilliant Blue (CBB) staining of the protein lysates of L. interrogans serovar Lai before (AP−) and after (AP+) alkaline phosphatase treating. (PDF 203 kb)
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Cao, XJ., Dai, J., Xu, H. et al. High-coverage proteome analysis reveals the first insight of protein modification systems in the pathogenic spirochete Leptospira interrogans. Cell Res 20, 197–210 (2010). https://doi.org/10.1038/cr.2009.127
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DOI: https://doi.org/10.1038/cr.2009.127
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