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Biotechniques. 2011 Jan;50(1):41-5. doi: 10.2144/000113586.

Bacterial flavohemoglobin: a molecular tool to probe mammalian nitric oxide biology.

Author information

  • 1Department of Biochemistry, Medical Scientist Training Program, Duke University Medical Center, Durham, NC 27710, USA. forrester@biochem.duke.edu

Abstract

A wide range of mammalian signaling and stress pathways are mediated by nitric oxide (NO), which is synthesized in vivo by the nitric oxide synthase (NOS) family of enzymes. Experimental manipulations of NO are frequently achieved by either inhibition or activation of endogenous NOS or via providing exogenous NO sources. On the contrary, many microbes consume NO via flavohemoglobin (FlavoHb), a highly efficient NO-dioxygenase that protects from nitrosative stress. Here we report a novel resource for studying NO in mammalian cells by heterologously expressing Escherichia coli FlavoHb within a lentiviral delivery system. This technique boosts endogenous cellular consumption of NO, thus providing a simple and efficacious approach to studying mammalian NO biology that can be employed as both a primary experimental and confirmatory tool.

PMID:
21231921
[PubMed - indexed for MEDLINE]
PMCID:
PMC3096140
Free PMC Article

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