Abstract:
:Apical caspases 8, 9, and 10 are only active as dimers. These dimers are unstable, and to characterize their activity they need to be maintained in vitro in a dimeric state. We provide updated methods for those looking to characterize various aspects of caspase function. We describe full methods for those looking to activate caspases in vitro using kosmotropic reagents, an essential step in characterizing upstream (apical) caspases. We detail methods for fusion of caspase domains to engineered dimerization domains as an alternative method to trigger regulated dimerization of caspases. We also describe methods to determine caspase activity profiles in cells and provide methods for studying the ability of SMAC-mimetic reagents to release inhibition of caspases by IAPs.
journal_name
Methods Enzymoljournal_title
Methods in enzymologyauthors
Mace PD,Riedl SJ,Salvesen GSdoi
10.1016/B978-0-12-417158-9.00007-8subject
Has Abstractpub_date
2014-01-01 00:00:00pages
161-78eissn
0076-6879issn
1557-7988pii
B978-0-12-417158-9.00007-8journal_volume
544pub_type
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