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Title: A constitutively active and uninhibitable caspase-3 zymogen efficiently induces apoptosis

Abstract

The caspase-3 zymogen has essentially zero activity until it is cleaved by initiator caspases during apoptosis. However, a mutation of V266E in the dimer interface activates the protease in the absence of chain cleavage. We show that low concentrations of the pseudo-activated procaspase-3 kill mammalian cells rapidly and, importantly, this protein is not cleaved nor is it inhibited efficiently by the endogenous regulator XIAP (X-linked inhibitor of apoptosis). The 1.63 {angstrom} (1 {angstrom} = 0.1 nm) structure of the variant demonstrates that the mutation is accommodated at the dimer interface to generate an enzyme with substantially the same activity and specificity as wild-type caspase-3. Structural modelling predicts that the interface mutation prevents the intersubunit linker from binding in the dimer interface, allowing the active sites to form in the procaspase in the absence of cleavage. The direct activation of procaspase-3 through a conformational switch rather than by chain cleavage may lead to novel therapeutic strategies for inducing cell death.

Authors:
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  1. (
Publication Date:
Research Org.:
Argonne National Lab. (ANL), Argonne, IL (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
1006109
Resource Type:
Journal Article
Resource Relation:
Journal Name: Biochem. J.; Journal Volume: 424; Journal Issue: 2009
Country of Publication:
United States
Language:
ENGLISH
Subject:
59 BASIC BIOLOGICAL SCIENCES; 99 GENERAL AND MISCELLANEOUS//MATHEMATICS, COMPUTING, AND INFORMATION SCIENCE; APOPTOSIS; CHAINS; CLEAVAGE; DEATH; DIMERS; ENZYMES; MUTATIONS; PROTEINS; SPECIFICITY

Citation Formats

Walters, Jad, Pop, Cristina, Scott, Fiona L., Drag, Marcin, Swartz, Paul, Mattos, Carla, Salvesen, Guy S., Clark, A.Clay, NCSU), and Burnham). A constitutively active and uninhibitable caspase-3 zymogen efficiently induces apoptosis. United States: N. p., 2010. Web. doi:10.1042/BJ20090825.
Walters, Jad, Pop, Cristina, Scott, Fiona L., Drag, Marcin, Swartz, Paul, Mattos, Carla, Salvesen, Guy S., Clark, A.Clay, NCSU), & Burnham). A constitutively active and uninhibitable caspase-3 zymogen efficiently induces apoptosis. United States. doi:10.1042/BJ20090825.
Walters, Jad, Pop, Cristina, Scott, Fiona L., Drag, Marcin, Swartz, Paul, Mattos, Carla, Salvesen, Guy S., Clark, A.Clay, NCSU), and Burnham). 2010. "A constitutively active and uninhibitable caspase-3 zymogen efficiently induces apoptosis". United States. doi:10.1042/BJ20090825.
@article{osti_1006109,
title = {A constitutively active and uninhibitable caspase-3 zymogen efficiently induces apoptosis},
author = {Walters, Jad and Pop, Cristina and Scott, Fiona L. and Drag, Marcin and Swartz, Paul and Mattos, Carla and Salvesen, Guy S. and Clark, A.Clay and NCSU) and Burnham)},
abstractNote = {The caspase-3 zymogen has essentially zero activity until it is cleaved by initiator caspases during apoptosis. However, a mutation of V266E in the dimer interface activates the protease in the absence of chain cleavage. We show that low concentrations of the pseudo-activated procaspase-3 kill mammalian cells rapidly and, importantly, this protein is not cleaved nor is it inhibited efficiently by the endogenous regulator XIAP (X-linked inhibitor of apoptosis). The 1.63 {angstrom} (1 {angstrom} = 0.1 nm) structure of the variant demonstrates that the mutation is accommodated at the dimer interface to generate an enzyme with substantially the same activity and specificity as wild-type caspase-3. Structural modelling predicts that the interface mutation prevents the intersubunit linker from binding in the dimer interface, allowing the active sites to form in the procaspase in the absence of cleavage. The direct activation of procaspase-3 through a conformational switch rather than by chain cleavage may lead to novel therapeutic strategies for inducing cell death.},
doi = {10.1042/BJ20090825},
journal = {Biochem. J.},
number = 2009,
volume = 424,
place = {United States},
year = 2010,
month = 3
}
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