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PMID: 16251271 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

Formation of apoptosome is initiated by cytochrome c-induced dATP hydrolysis and subsequent nucleotide exchange on Apaf-1.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 102 ·No. 49 ·2005-12-06 ·Pages 17545-50

Kim HE, Du F, Fang M, Wang X

Abstract

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.

MeSH Terms
Animals Apoptosis Apoptotic Protease-Activating Factor 1 Caspase 3 Caspases/metabolism Cytochromes c/metabolism Deoxyadenine Nucleotides/metabolism Enzyme Activation Horses Hydrolysis Intracellular Membranes/metabolism Intracellular Signaling Peptides and Proteins/genetics,metabolism Organelles/metabolism Proteins/genetics,metabolism
Chemicals
Apoptotic Protease-Activating Factor 1 Deoxyadenine Nucleotides Intracellular Signaling Peptides and Proteins Proteins Cytochromes c Caspase 3 Caspases 2'-deoxyadenosine triphosphate
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Kim Hyun-Eui
Howard Hughes Medical Institute and Department of Biochemistry, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA.
Du Fenghe
Fang Min
Wang Xiaodong
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.0507900102
Published
2005-12-06
Epub
2005-00-26
Pages
17545-50
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC1266161
Subset
IM
Grants
NIGMS NIH HHS · GMR01 57158 · United States
External Links
PubMed source DOI: 10.1073/pnas.0507900102 PMC Full Text View journal details
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