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Kinetic flux profiling of nitrogen assimilation in Escherichia coli

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Abstract

We present a new method for probing cellular metabolic fluxes that is based on the kinetics of assimilation of isotope-labeled nutrient into a diversity of downstream metabolites. In the case of nitrogen assimilation, half-maximal labeling of most metabolites occurs in 10–300 s. Fluxes measured on the basis of the kinetics of nitrogen assimilation in exponentially growing E. coli agree well with those fluxes predicted to allow optimal biomass production.

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Figure 1: Kinetics of isotope labeling of the central intermediates in nitrogen assimilation.

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References

  1. Schilling, C.H. & Palsson, B.O. Proc. Natl. Acad. Sci. USA 95, 4193–4198 (1998).

    Article  CAS  Google Scholar 

  2. Edwards, J.S., Ibarra, R.U. & Palsson, B.O. Nat. Biotechnol. 19, 125–130 (2001).

    Article  CAS  Google Scholar 

  3. Stephanopoulos, G. Metab. Eng. 1, 1–11 (1999).

    Article  CAS  Google Scholar 

  4. Szyperski, T. Eur. J. Biochem. 232, 433–448 (1995).

    Article  CAS  Google Scholar 

  5. Chatham, J.C., Forder, J.R., Glickson, J.D. & Chance, E.M. J. Biol. Chem. 270, 7999–8008 (1995).

    Article  CAS  Google Scholar 

  6. Fischer, E. & Sauer, U. Eur. J. Biochem. 270, 880–891 (2003).

    Article  CAS  Google Scholar 

  7. van Winden, W.A. et al. FEMS Yeast Res. 5, 559–568 (2005).

    Article  CAS  Google Scholar 

  8. Fischer, E. & Sauer, U. Nat. Genet. 37, 636–640 (2005).

    Article  CAS  Google Scholar 

  9. Ibarra, R.U., Edwards, J.S. & Palsson, B.Ø. Nature 420, 186–189 (2002).

    Article  CAS  Google Scholar 

  10. Segre, D., Vitkup, D. & Church, G.M. Proc. Natl. Acad. Sci. USA 99, 15112–15117 (2002).

    Article  CAS  Google Scholar 

  11. Lu, W., Kimball, E. & Rabinowitz, J.D. J. Am. Soc. Mass Spectrom. 17, 37–50 (2006).

    Article  CAS  Google Scholar 

  12. Ikeda, T.P., Shauger, A.E. & Kustu, S. J. Mol. Biol. 259, 589–607 (1996).

    Article  CAS  Google Scholar 

  13. Reitzer, L. Annu. Rev. Microbiol. 57, 155–176 (2003).

    Article  CAS  Google Scholar 

  14. Helling, R.B. J. Bacteriol. 184, 1041–1045 (2002).

    Article  CAS  Google Scholar 

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Acknowledgements

We thank D. Botstein and N. Wingreen for their intellectual contributions and the US National Science Foundation, the National Institutes of Health and the Beckman Foundation for their financial support.

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Authors and Affiliations

  1. Department of Chemistry and Lewis-Sigler Institute for Integrative Genomics, Princeton University, Princeton, 08544, New Jersey, USA

    Jie Yuan, William U Fowler, Elizabeth Kimball, Wenyun Lu & Joshua D Rabinowitz

Authors
  1. Jie Yuan
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  2. William U Fowler
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  3. Elizabeth Kimball
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  4. Wenyun Lu
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  5. Joshua D Rabinowitz
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Corresponding author

Correspondence to Joshua D Rabinowitz.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Fig. 1

Passage of isotope-labeled nitrogen from central assimilation intermediates into biopolymer precursors. (PDF 45 kb)

Supplementary Fig. 2

Effect of carbon starvation on intracellular metabolite pools and fluxes in E. coli. (PDF 39 kb)

Supplementary Methods (PDF 66 kb)

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Yuan, J., Fowler, W., Kimball, E. et al. Kinetic flux profiling of nitrogen assimilation in Escherichia coli. Nat Chem Biol 2, 529–530 (2006). https://doi.org/10.1038/nchembio816

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