TY - JOUR
T1 - Cytochrome c and dATP-dependent formation of Apaf-1/caspase-9 complex initiates an apoptotic protease cascade
AU - Li, Peng
AU - Nijhawan, Deepak
AU - Budihardjo, Imawati
AU - Srinivasula, Srinivasa M.
AU - Ahmad, Manzoor
AU - Alnemri, Emad S.
AU - Wang, Xiaodong
N1 - Funding Information:
We thank our colleagues Xuesong Liu, Hua Zou, Mike Lutter, and Holt Oliver for helpful discussions and suggestions, and Alexis Lutschg for excellent technical assistance. We thank Dr. Clive Slaughter, Carolyn Moomaw, and Steve Afendis for help with the protein sequencing analysis. We also thank Dr. Marion Macfarlane for help in the apoptosis assays. We are grateful to our colleagues Drs. Joseph Goldstein, Michael Brown, and Teresa Fernandes-Alnemri for critically reading the manuscript. Suspension cultured HeLa cells were obtained from the Cell Culture Center at Minneapolis. D. N. is supported by Medical Scientists Training Grant GM08014. X. W. is also supported by an American Cancer Society Research Grant and a NIH GMRO1–55942. This work was also supported in part by research grant AG13487 from the NIH (E. S. A).
PY - 1997/11/14
Y1 - 1997/11/14
N2 - We report here the purification of the third protein factor, Apaf-3, that participates in caspase-3 activation in vitro. Apaf-3 was identified as a member of the caspase family, caspase-9. Caspase-9 and Apaf-1 bind to each other via their respective NH2-terminal CED-3 homologous domains in the presence of cytochrome c and dATP, an event that leads to caspase-9 activation. Activated caspase-9 in turn cleaves and activates caspase-3. Depletion of caspase-9 from S-100 extracts diminished caspase-3 activation. Mutation of the active site of caspase-9 attenuated the activation of caspase-3 and cellular apoptotic response in vivo, indicating that caspase-9 is the most upstream member of the apoptotic protease cascade that is triggered by cytochrome c and dATP.
AB - We report here the purification of the third protein factor, Apaf-3, that participates in caspase-3 activation in vitro. Apaf-3 was identified as a member of the caspase family, caspase-9. Caspase-9 and Apaf-1 bind to each other via their respective NH2-terminal CED-3 homologous domains in the presence of cytochrome c and dATP, an event that leads to caspase-9 activation. Activated caspase-9 in turn cleaves and activates caspase-3. Depletion of caspase-9 from S-100 extracts diminished caspase-3 activation. Mutation of the active site of caspase-9 attenuated the activation of caspase-3 and cellular apoptotic response in vivo, indicating that caspase-9 is the most upstream member of the apoptotic protease cascade that is triggered by cytochrome c and dATP.
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U2 - 10.1016/S0092-8674(00)80434-1
DO - 10.1016/S0092-8674(00)80434-1
M3 - Article
C2 - 9390557
AN - SCOPUS:0030715323
SN - 0092-8674
VL - 91
SP - 479
EP - 489
JO - Cell
JF - Cell
IS - 4
ER -