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J. Biol. Chem., Vol. 276, Issue 5, 3098-3105, February 2, 2001
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From the Section on Metabolic Regulation, Endocrinology and
Reproduction Research Branch, NICHD, National Institutes of Health,
Bethesda, Maryland 20892-4510
S-Nitrosoglutathione (GSNO) undergoes
spontaneous degradation that generates several nitrogen-containing
compounds and oxidized glutathione derivatives. We identified
glutathione sulfonic acid, glutathione disulfide
S-oxide (GS(O)SG), glutathione disulfide S-dioxide, and GSSG as the major decomposition
products of GSNO. Each of these compounds and GSNO were tested for
their efficacies to modify rat brain neurogranin/RC3 (Ng) and
neuromodulin/GAP-43 (Nm). Among them, GS(O)SG was found to be the most
potent in causing glutathiolation of both proteins; four
glutathiones were incorporated into the four Cys residues of Ng,
and two were incorporated into the two Cys residues of Nm. Ng and Nm
are two in vivo substrates of protein kinase C; their
phosphorylations by protein kinase C attenuate the binding affinities
of both proteins for calmodulin. When compared with their respective
unmodified forms, the glutathiolated Ng was a poorer substrate and
glutathiolated Nm a better substrate for protein kinase C. Glutathiolation of these two proteins caused no change in their binding
affinities for calmodulin. Treatment of
[35S]cysteine-labeled rat brain slices with
xanthine/xanthine oxidase or a combination of xanthine/xanthine oxidase
with sodium nitroprusside resulted in an increase in cellular level of
GS(O)SG. These treatments, as well as those by other oxidants, all
resulted in an increase in thiolation of proteins; among them,
thiolation of Ng was positively identified by immunoprecipitation.
These results show that GS(O)SG is one of the most potent
glutathiolating agents generated upon oxidative stress.
To whom correspondence should be addressed: Bldg. 49, Rm. 6A36,
NIH, 49 Convent Dr., MSC 4510, Bethesda, MD 20892-4510. Tel.: 301-496-7827; Fax: 301-496-7434; E-mail kphuang@helix.nih.gov.
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