Quantifying Nitric Oxide Flux Distributions
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Nitric oxide (NO) is a radical that is used as an attack molecule by immune cells. NO can interact and damage a range of biomolecules, and the biological outcome for bacteria assaulted with NO will be governed by how the radical distributes within their biochemical reaction networks. Measurement of those NO fluxes is complicated by the low abundance and transience of many of its reaction products. To overcome this challenge, we use computational modeling to translate measurements of several biochemical species (e.g., NO, O2, NO2−) into NO flux distributions. In this chapter, we provide a detailed protocol, which includes experimental measurements and computational modeling, to estimate the NO flux distribution in an Escherichia coli culture. Those fluxes will have uncertainty associated with them and we also discuss how further experiments and modeling can be employed for flux refinement.
Key wordsEscherichia coli Nitric oxide Metabolic flux Nitric oxide dioxygenase Nitric oxide reductase
This work was supported by National Science Foundation grant CBET-1453325 (MPB), Natural Sciences and Engineering Research Council of Canada Postgraduate Scholarship (DMS), and the generosity of Helen Shipley Hunt ∗71 through the Shipley Hunt Fund (MPB). We would like to thank Weng Kang Chou and Annabel S. Lemma for their assistance. The funders had no role in the preparation of the manuscript or decision to publish, and this content is solely the responsibility of the authors and does not necessarily represent the views of the funding agencies.
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