Identification of a denitrase activity against calmodulin in activated macrophages using high-field liquid chromatography-FTICR mass spectrometry

Heather Smallwood, Natacha M. Lourette, Curt B. Boschek, Diana J. Bigelow, Richard D. Smith, Ljiljana Paša-Tolić, Thomas C. Squier

Research output: Contribution to journalArticle

46 Citations (Scopus)

Abstract

We have identified a denitrase activity in macrophages that is upregulated following macrophage activation, which is shown by mass spectrometry to recognize nitrotyrosines in the calcium signaling protein calmodulin (CaM). The denitrase activity converts nitrotyrosines to their native tyrosine structure without the formation of any aminotyrosine. Comparable extents of methionine sulfoxide reduction are also observed that are catalyzed by endogenous methionine sulfoxide reductases. Competing with repair processes, oxidized CaM is a substrate for a peptidase activity that results in the selective cleavage of the C-terminal lysine (i.e., Lys148) that is expected to diminish CaM function. Thus, competing repair and peptidase activities define the abundances and functionality of CaM in modulating cellular metabolism in response to oxidative stress, where the presence of the truncated CaM species provides a useful biomarker for the transient appearance of oxidized CaM.

Original languageEnglish (US)
Pages (from-to)10498-10505
Number of pages8
JournalBiochemistry
Volume46
Issue number37
DOIs
StatePublished - Sep 18 2007
Externally publishedYes

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Macrophages
Liquid chromatography
Calmodulin
Liquid Chromatography
Mass spectrometry
Mass Spectrometry
Methionine Sulfoxide Reductases
Repair
Peptide Hydrolases
Oxidative stress
Calcium Signaling
Macrophage Activation
Biomarkers
Metabolism
Lysine
Tyrosine
Oxidative Stress
Chemical activation
Calcium
Substrates

All Science Journal Classification (ASJC) codes

  • Biochemistry

Cite this

Identification of a denitrase activity against calmodulin in activated macrophages using high-field liquid chromatography-FTICR mass spectrometry. / Smallwood, Heather; Lourette, Natacha M.; Boschek, Curt B.; Bigelow, Diana J.; Smith, Richard D.; Paša-Tolić, Ljiljana; Squier, Thomas C.

In: Biochemistry, Vol. 46, No. 37, 18.09.2007, p. 10498-10505.

Research output: Contribution to journalArticle

Smallwood, Heather ; Lourette, Natacha M. ; Boschek, Curt B. ; Bigelow, Diana J. ; Smith, Richard D. ; Paša-Tolić, Ljiljana ; Squier, Thomas C. / Identification of a denitrase activity against calmodulin in activated macrophages using high-field liquid chromatography-FTICR mass spectrometry. In: Biochemistry. 2007 ; Vol. 46, No. 37. pp. 10498-10505.
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AU - Paša-Tolić, Ljiljana

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