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J. Biol. Chem., Vol. 277, Issue 5, 3614-3621, February 1, 2002
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From the Nitric oxide-derived reactive species have been
implicated in many disorders. Protein nitrotyrosine is often used as a
stable marker of these reactive species. Using immunohistochemistry, we
have previously detected nitrotyrosine in murine Mutatect tumors, where
neutrophils are the principal source of nitric oxide. We now report on
the identification of several prominent nitrotyrosine-containing proteins. Using Western blot analysis, nitrotyrosine in higher molecular mass proteins (>20 kDa) was detected in tumors containing a
high number of neutrophils but not in tumors with fewer neutrophils. Staining for nitrotyrosine was consistently seen in low molecular mass
proteins (
Selective Nitration of Histone Tyrosine Residues in
Vivo in Mutatect Tumors*
§,
Department of Biochemistry, Microbiology and
Immunology, University of Ottawa and the Ottawa Regional Cancer Centre,
Ottawa, Ontario K1H 1C4, Canada and the ¶ Institute of Biological
Sciences, 100 Sussex Drive, Ottawa, Ontario K1A 0R6, Canada
15 kDa), regardless of the level of neutrophils. Protein
nitrotyrosine was not seen in Mutatect cells growing in vitro. Treatment with nitric oxide donors produced nitration of 15-kDa proteins, but only after extended periods. These small proteins, both from tumors and cultured cells, were identified by mass
spectrometry to be histones. Only a subset of tyrosine residues was
nitrated. Selective nitration may reflect differential accessibility of
different tyrosine residues and the influence of neighboring residues
within the nucleosome. The prominence of histone nitration may reflect
its relative stability, making this post-translational modification a
potentially useful marker of extended exposure of cells or tissues to
nitric oxide-derived reactive species.
*
This work was supported in part by grants from the Cancer
Research Society and Canadian Institutes of Health Research (to H. C. B.).The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
Senior Scientist of the Ottawa Regional Cancer Center. To whom
correspondence should be addressed: Ottawa Regional Cancer Center, 503 Smyth Rd., Ottawa, Ontario K1H 1C4, Canada. Tel.: 613-737-7700 (ext.
6701); Fax: 613-247-3524; E-mail: birnboim@uottawa.ca.
Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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