Solution structure and mutagenesis of the caspase recruitment domain (CARD) from Apaf-1.
Activation of procaspase-9, a key component of the apoptosis mechanism, requires the interaction of its caspase recruitment domain (CARD) with the CARD in the adaptor protein Apaf-1. Using nuclear magnetic resonance spectroscopy and mutagenesis we have determined the structure of the CARD from Apaf-1 and the residues important for binding the CARD in procaspase-9. Apaf-1's CARD contains seven short alpha-helices with the core six helices arranged in an antiparallel manner. Residues in helix 2 have a central role in mediating interaction with procaspase-9 CARD. This interaction surface is distinct from that proposed based on the structure of the CARD from RAIDD, but is coincident with that of the structurally similar FADD death effector domain and the Apaf-1 CARD interface identified by crystallographic studies.
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Institute of Molecular BioSciences, Massey University, Private Bag 11 222, Palmerston North, New Zealand.
Dupont
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Lackmann
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Vaux
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DL
Hinds
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MG
D048868:Adaptor Proteins, Signal Transducing; D000595:Amino Acid Sequence; D053477:Apoptotic Protease-Activating Factor 1; D001665:Binding Sites; D002352:Carrier Proteins / Q000737:chemistry; D053453:Caspase 9; D020169:Caspases / Q000737:chemistry / Q000378:metabolism*; D053200:Fas-Associated Death Domain Protein; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016296:Mutagenesis; D011487:Protein Conformation; D011506:Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012996:Solutions
C506592:APAF1 protein, human; D048868:Adaptor Proteins, Signal Transducing; D053477:Apoptotic Protease-Activating Factor 1; D002352:Carrier Proteins; C505533:FADD protein, human; D053200:Fas-Associated Death Domain Protein; D011506:Proteins; D011993:Recombinant Fusion Proteins; D012996:Solutions; C506433:CASP9 protein, human; D053453:Caspase 9; D020169:Caspases