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TitleMechanistic insights into caspase-9 activation by the structure of the apoptosome holoenzyme.
Journal, issue, pagesProc Natl Acad Sci U S A, Vol. 114, Issue 7, Page 1542-1547, Year 2017
Publish dateFeb 14, 2017
AuthorsYini Li / Mengying Zhou / Qi Hu / Xiao-Chen Bai / Weiyun Huang / Sjors H W Scheres / Yigong Shi /
PubMed AbstractMammalian intrinsic apoptosis requires activation of the initiator caspase-9, which then cleaves and activates the effector caspases to execute cell killing. The heptameric Apaf-1 apoptosome is ...Mammalian intrinsic apoptosis requires activation of the initiator caspase-9, which then cleaves and activates the effector caspases to execute cell killing. The heptameric Apaf-1 apoptosome is indispensable for caspase-9 activation by together forming a holoenzyme. The molecular mechanism of caspase-9 activation remains largely enigmatic. Here, we report the cryoelectron microscopy (cryo-EM) structure of an apoptotic holoenzyme and structure-guided biochemical analyses. The caspase recruitment domains (CARDs) of Apaf-1 and caspase-9 assemble in two different ways: a 4:4 complex docks onto the central hub of the apoptosome, and a 2:1 complex binds the periphery of the central hub. The interface between the CARD complex and the central hub is required for caspase-9 activation within the holoenzyme. Unexpectedly, the CARD of free caspase-9 strongly inhibits its proteolytic activity. These structural and biochemical findings demonstrate that the apoptosome activates caspase-9 at least in part through sequestration of the inhibitory CARD domain.
External linksProc Natl Acad Sci U S A / PubMed:28143931 / PubMed Central
MethodsEM (single particle)
Resolution4.4 - 6.9 Å
Structure data

6690

5wve

EMDB-6690, PDB-5wve:
Apaf-1-Caspase-9 holoenzyme
Method: EM (single particle) / Resolution: 4.4 Å

EMDB-6691:
Apaf-1-Caspase-9 holoenzyme
Method: EM (single particle) / Resolution: 4.4 Å

EMDB-6692:
Apaf-1-Caspase-9 holoenzyme
Method: EM (single particle) / Resolution: 6.9 Å

Chemicals

ChemComp-DTP:
2'-DEOXYADENOSINE 5'-TRIPHOSPHATE / Deoxyadenosine triphosphate

ChemComp-MG:
Unknown entry

ChemComp-HEM:
PROTOPORPHYRIN IX CONTAINING FE / Heme B

Source
  • homo sapiens (human)
  • equus caballus (horse)
KeywordsAPOPTOSIS / apoptosis holoenzyme

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