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J. Biol. Chem., Vol. 275, Issue 31, 24136-24145, August 4, 2000
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From the We investigated the effects of mild oxidation on
protein kinase C (PKC) using the xanthine/xanthine oxidase system of
generating superoxide. Exposure of various PKC preparations to
superoxide stimulated the autonomous activity of PKC. Similarly, thiol
oxidation increased autonomous PKC activity, consistent with the notion that superoxide stimulates PKC via thiol oxidation. The
superoxide-induced stimulation of PKC activity was partially reversed
by reducing agents, suggesting that disulfide bond formation
contributed to the oxidative stimulation of PKC. In addition,
superoxide increased the autonomous activity of the
Superoxide-induced Stimulation of Protein Kinase C via Thiol
Modification and Modulation of Zinc Content*
and§¶
Department of Neuroscience and the
§ Center for the Neural Basis of Cognition, University of
Pittsburgh, Pittsburgh, Pennsylvania 15260
,
II, 
, and 
PKC isoforms, all of which
contain at least one cysteine-rich region. Taken together, our
observations suggested that superoxide interacts with PKC at the
cysteine-rich region, zinc finger motif of the enzyme. Therefore, we
examined the effects of superoxide on this region by testing the
hypothesis that superoxide stimulates PKC by promoting the release of
zinc from PKC. We found that a zinc chelator stimulated the autonomous
activity of PKC and that superoxide induced zinc release from an
PKC-enriched enzyme preparation. In addition, oxidized PKC contained
significantly less zinc than reduced PKC. Finally, we have isolated a
persistent, autonomously active PKC by DEAE-cellulose column
chromatography from hippocampal slices incubated with superoxide. Taken
together, these data suggest that superoxide stimulates autonomous PKC
activity via thiol oxidation and release of zinc from cysteine-rich
region of PKC.
*
This work was supported by National Institutes of Health
Grants NS08950 and NS34007 (both to E. K.), National Institute of Mental Health Training Grant MH18273, National Institute of Mental Health National Research Service Award Fellowship MH1198301 (to L. T. K.), a University of Pittsburgh Central Research Development Fund award (to E. K.), and a grant from the Winters
Foundation (to E. K.).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.
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