Premium
Heterologous expression of microbial flavohemoglobin can modulate the effects of nitric oxide in mammalian cells
Author(s) -
Eyler Christine Elissa,
Forrester Michael T.,
Hjelmeland Anita B.,
Rich Jeremy N.
Publication year - 2010
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.24.1_supplement.871.2
Subject(s) - intracellular , nitric oxide , microbiology and biotechnology , heterologous , nitric oxide synthase , signal transduction , heterologous expression , biology , reactive nitrogen species , enzyme , reactive oxygen species , biochemistry , chemistry , gene , recombinant dna , endocrinology
Nitric oxide (NO) contributes to a wide variety of intracellular signaling and cell stress pathways in mammalian cells. Experimental manipulation of NO is frequently achieved by inhibition or activation of the NO synthase (NOS) family of enzymes responsible for intracellular NO production, or through employment of relatively nonspecific NO‐reactive compounds or antioxidants. Here we introduce a novel methodology to manipulate NO levels in mammalian systems via heterologous expression of the prokaryotic NO‐consuming flavohemoglobin (FlavoHb). This enzyme is an NO‐specific dioxygenase that converts NO to inert nitrate under aerobic conditions, thus functioning to consume NO and protect microbes from nitrosative stress. We demonstrate that FlavoHb can be employed in a similar fashion in mammalian cells to inhibit both NO‐dependent signal transduction and nitrosative stress. This strategy provides a highly specific and novel method for modulating intracellular NO levels and represents an alternative or complimentary strategy to utilization of NOS inhibitors or promiscuous antioxidants. Importantly, FlavoHb expression is well tolerated by mammalian cells with no evidence of NO‐independent toxicity. Future studies of NO in mammalian systems could employ FlavoHb as either a primary experimental or a confirmatory tool for probing functional and molecular consequences of NO within cells.