Nitric oxide dioxygenation reaction in devS and the initial response to nitric oxide in mycobacterium tuberculosis

Erik T. Yukl, Alexandra Ioanoviciu, Santhosh Sivaramakrishnan, Michiko Nakano, Paul R. Ortiz De Montellano, Pierre Moenne-Loccoz

Research output: Contribution to journalArticle

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Abstract

DevS and DosT from Mycobacterium tuberculosis (MTB) are paralogous heme-based sensor kinases that respond to hypoxia and to low concentrations of nitric oxide (NO). Both proteins work with the response regulator DevR as a two-component regulatory system to induce the dormancy regulon in MTB. While DevS and DosT are inactive when dioxygen is bound to the heme Fe(II) at their sensor domain, autokinase activity is observed in their heme Fe(II)-NO counterparts. To date, the conversion between active and inactive states and the reactivity of the heme-oxy complex toward NO have not been investigated. Here, we use stopped-flow UV-vis spectroscopy and rapid freeze quench resonance Raman spectroscopy to probe these reactions in DevS. Our data reveal that the heme-O2 complex of DevS reacts efficiently with NO to produce nitrate and the oxidized Fe(III) heme through an NO dioxygenation reaction that parallels the catalytic reactions of bacterial flavohemoglobin and truncated hemoglobins. Autophosphorylation activity assays show that the Fe(III) heme state of DevS remains inactive but exhibits a high affinity for NO and forms an Fe(III)-NO complex that is readily reduced by ascorbate, a mild reducing agent. On the basis of these results, we conclude that upon exposure to low NO concentrations, the inactive oxy-heme complex of DevS is rapidly converted to the Fe(II)-NO complex in the reducing environment of living cells and triggers the initiation of dormancy.

Original languageEnglish (US)
Pages (from-to)1023-1028
Number of pages6
JournalBiochemistry
Volume50
Issue number6
DOIs
StatePublished - Feb 15 2011

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Mycobacterium tuberculosis
Heme
Nitric Oxide
Truncated Hemoglobins
Regulon
Raman Spectrum Analysis
Reducing Agents
Sensors
Ultraviolet spectroscopy
Nitrates
Raman spectroscopy
Assays
Spectrum Analysis
Phosphotransferases
Cells
Oxygen

ASJC Scopus subject areas

  • Biochemistry

Cite this

Nitric oxide dioxygenation reaction in devS and the initial response to nitric oxide in mycobacterium tuberculosis. / Yukl, Erik T.; Ioanoviciu, Alexandra; Sivaramakrishnan, Santhosh; Nakano, Michiko; Ortiz De Montellano, Paul R.; Moenne-Loccoz, Pierre.

In: Biochemistry, Vol. 50, No. 6, 15.02.2011, p. 1023-1028.

Research output: Contribution to journalArticle

Yukl, Erik T. ; Ioanoviciu, Alexandra ; Sivaramakrishnan, Santhosh ; Nakano, Michiko ; Ortiz De Montellano, Paul R. ; Moenne-Loccoz, Pierre. / Nitric oxide dioxygenation reaction in devS and the initial response to nitric oxide in mycobacterium tuberculosis. In: Biochemistry. 2011 ; Vol. 50, No. 6. pp. 1023-1028.
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