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Briefings in Functional Genomics and Proteomics Advance Access originally published online on May 23, 2006
Briefings in Functional Genomics and Proteomics 2006 5(2):133-143; doi:10.1093/bfgp/ell025
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© Oxford University Press, 2006, All rights reserved. For permissions, please email: journals.permissions@oxfordjournals.org

Special Issue Papers

Study of nitrate stress in Desulfovibrio vulgaris Hildenborough using iTRAQ proteomics

Alyssa M. Redding, Aindrila Mukhopadhyay, Dominique C. Joyner, Terry C. Hazen and Jay D. Keasling

Corresponding author. Prof. Jay D. Keasling, Berkeley Center for Synthetic Biology, University of California, Berkeley, 717 Potter St., Bldg 977 MC 3224, Berkeley, CA 94720-3224 USA. Tel: 510-495-2620; Fax: 510-495-2630; E-mail: keasling{at}berkeley.edu

The response of Desulfovibrio vulgaris Hildenborough (DvH), a sulphate-reducing bacterium, to nitrate stress was examined using quantitative proteomic analysis. DvH was stressed with 105 mM sodium nitrate (NaNO3), a level that caused a 50% inhibition in growth. The protein profile of stressed cells was compared with that of cells grown in the absence of nitrate using the iTRAQ peptide labelling strategy and tandem liquid chromatography separation coupled with mass spectrometry (quadrupole time-of-flight) detection. A total of 737 unique proteins were identified by two or more peptides, representing 22% of the total DvH proteome and spanning every functional category. The results indicate that this was a mild stress, as proteins involved in central metabolism and the sulphate reduction pathway were unperturbed. Proteins involved in the nitrate reduction pathway increased. Increases seen in transport systems for proline, glycine–betaine and glutamate indicate that the NaNO3 exposure led to both salt stress and nitrate stress. Up-regulation observed in oxidative stress response proteins (Rbr, RbO, etc.) and a large number of ABC transport systems as well as in iron–sulphur-cluster-containing proteins, however, appear to be specific to nitrate exposure. Finally, a number of hypothetical proteins were among the most significant changers, indicating that there may be unknown mechanisms initiated upon nitrate stress in DvH.

Keywords: Desulfovibro vulgaris, iTRAQ, peptide tagging, nitrate


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