Formation of Apoptosome Is Initiated by Cytochrome c-Induced dATP Hydrolysis and Subsequent Nucleotide Exchange on Apaf-1

Apoptosis in metazoans is executed by a group of intracellular proteases named caspases. One of the caspase-activating pathways in mammals is initiated by the release of cytochrome c from mitochondria to cytosol, where it binds to Apaf-1 to form a procaspase-9-activating heptameric protein complex n...

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Published inProceedings of the National Academy of Sciences - PNAS Vol. 102; no. 49; pp. 17545 - 17550
Main Authors Kim, Hyun-Eui, Du, Fenghe, Fang, Min, Wang, Xiaodong
Format Journal Article
LanguageEnglish
Published United States National Academy of Sciences 06.12.2005
National Acad Sciences
SeriesInaugural Article
Subjects
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Summary:Apoptosis in metazoans is executed by a group of intracellular proteases named caspases. One of the caspase-activating pathways in mammals is initiated by the release of cytochrome c from mitochondria to cytosol, where it binds to Apaf-1 to form a procaspase-9-activating heptameric protein complex named apoptosome. We report here the reconstitution of this pathway with purified recombinant Apaf-1, procaspase-9, procaspase-3, and cytochrome c from horse heart. Apaf-1 contains a dATP as a cofactor. Cytochrome c binding to Apaf-1 induces hydrolysis of dATP to dADP, which is subsequently replaced by exogenous dATP. The dATP hydrolysis and exchange on Apaf-1 are two required steps for apoptosome formation.
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To whom correspondence should be addressed. E-mail: xwang@biochem.swmed.edu.
This contribution is part of the special series of Inaugural Articles by members of the National Academy of Sciences elected on April 20, 2004.
Conflict of interest statement: No conflicts declared.
Contributed by Xiaodong Wang, September 13, 2005
Author contributions: H.-E.K., F.D., M.F., and X.W. designed research; H.-E.K., F.D., and M.F. performed research; F.D. contributed new reagents/analytic tools; H.-E.K., M.F., and X.W. analyzed data; and H.-E.K. and X.W. wrote the paper.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.0507900102