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J Biol Chem, Vol. 273, Issue 14, 7787-7790, April 3, 1998
From the Burnham Institute, Program on Apoptosis & Cell Death
Regulation, La Jolla, California 92037
The inhibitor of apoptosis proteins (IAPs)
constitute an evolutionarily conserved family of homologous proteins
that suppress apoptosis induced by multiple stimuli. Some IAP family
proteins, including XIAP, cIAP-1, and cIAP-2, can bind and directly
inhibit selected caspases, a group of intracellular cell death
proteases. These caspase-inhibiting IAP family proteins all contain
three tandem BIR domains followed by a RING zinc finger domain. To
determine the structural basis for caspase inhibition by XIAP, we
analyzed the effects of various fragments of this IAP family protein on caspase activity in vitro and on apoptosis suppression in
intact cells. The RING domain of XIAP failed to inhibit the activity of
recombinant caspases-3 or -7, whereas a fragment of XIAP encompassing the three tandem BIR domains potently inhibited these caspases in
vitro and blocked Fas (CD95)-induced apoptosis when expressed in
cells. Further dissection of the XIAP protein demonstrated that only
the second of the three BIR domains (BIR2) was capable of binding and
inhibiting these caspases. The apparent inhibition constants
(Ki) for BIR2-mediated inhibition of caspases-3 and
-7 were 2-5 nM, indicating that this single BIR domain
possesses potent anti-caspase activity. Expression of the BIR2 domain
in cells also partially suppressed Fas-induced apoptosis and
blocked cytochrome c-induced processing of caspase-9 in
cytosolic extracts, whereas BIR1 and BIR3 did not. These findings
identify BIR2 as the minimal caspase-inhibitory domain of XIAP and
indicate that a single BIR domain can be sufficient for binding and
inhibiting caspases.
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