Caspase enzymology and activation mechanisms.

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 Enzymol

journal_title

Methods in enzymology

authors

Mace PD,Riedl SJ,Salvesen GS

doi

10.1016/B978-0-12-417158-9.00007-8

subject

Has Abstract

pub_date

2014-01-01 00:00:00

pages

161-78

eissn

0076-6879

issn

1557-7988

pii

B978-0-12-417158-9.00007-8

journal_volume

544

pub_type

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