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J. Biol. Chem., Vol. 277, Issue 41, 38079-38086, October 11, 2002
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From the Department of Chemistry, University of Minnesota,
Duluth, Minnesota 55812
We reported that the generation
of nitric oxide by mitochondria is catalyzed by a constitutive,
mitochondrial nitric-oxide synthase (mtNOS). Given that this production
may establish the basis for a novel regulatory pathway of energy
metabolism, oxygen consumption, and oxygen free radical production, it
becomes imperative to identify unequivocally and characterize this
enzyme to provide a basis for its regulation. The mitochondrial
localization of mtNOS was supported by following the hepatic
distribution of mtNOS, immunoblotting submitochondrial fractions, and
immunohistochemistry of liver tissues. mtNOS was identified as brain
NOS
Biochemistry of Mitochondrial Nitric-oxide Synthase*
by various methods (mass spectrometry of proteolytic fragments,
amino acid analysis, molecular weight, pI, and analysis of PCR
fragments), excluding the occurrence of a novel isoform or other splice
variants. Distribution of mtNOS transcript indicated its occurrence in
liver, brain, heart, muscle, kidney, lung, testis, and spleen. In
contrast to brain NOS, mtNOS has two post-translational modifications: acylation with myristic acid and phosphorylation at the C terminus. The
former modification is a reversible and post-translational process,
which may serve for subcellular targeting or membrane anchoring. The
latter modification could be linked to enzymatic regulation. These
results are discussed in terms of the role that nitric oxide may have
in cellular bioenergetics.
*
This work was supported by National Institutes of Health
Grant ES011407-01, Minnesota Medical Foundation Grant 3048-9227-01, the
University of Minnesota Graduate School United Mitochondrial Disease
Foundation, and Cottrell Research Corporation Grant CC5675.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.
To whom correspondence should be addressed: Dept. of Chemistry,
University of Minnesota, 10 University Dr., Duluth, MN 55812. E-mail: cgiulivi@d.umn.edu.
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