Structure of an apoptosome-procaspase-9 CARD complex.
Apaf-1 coassembles with cytochrome c to form the apoptosome, which then binds and activates procaspase-9 (pc-9). We removed pc-9 catalytic domains from the holoapoptosome by site-directed thrombinolysis. A structure of the resulting apoptosome-pc-9 CARD complex was then determined at approximately 9.5 A resolution. In our model, the central hub is constructed like other AAA+ protein rings but also contains novel features. At higher radius, the regulatory region of each Apaf-1 is comprised of tandem seven and eight blade beta-propellers with cytochrome c docked between them. Remarkably, Apaf-1 CARDs are disordered in the ground state. During activation, each Apaf-1 CARD interacts with a pc-9 CARD and these heterodimers form a flexibly tethered "disk" that sits above the central hub. When taken together, the data reveal conformational changes during Apaf-1 assembly that allow pc-9 activation. The model also provides a plausible explanation for the effects of NOD mutations that have been mapped onto the central hub.
Yuan
Shujun
S
Department of Physiology and Biophysics, Boston University School of Medicine, 700 Albany Street, Boston, MA 02118-2526, USA.
Yu
Xinchao
X
Topf
Maya
M
Ludtke
Steven J
SJ
Wang
Xiaodong
X
Akey
Christopher W
CW
D053478:Apoptosomes / Q000378:metabolism*; D053453:Caspase 9 / Q000378:metabolism*; D003574:Cytochrome c Group; D045304:Cytochromes c / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D006801:Humans; D011506:Proteins / Q000737:chemistry* / Q000378:metabolism* / Q000502:physiology
D053478:Apoptosomes; D003574:Cytochrome c Group; D011506:Proteins; C076615:cytochrome c''; D045304:Cytochromes c; D053453:Caspase 9
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