Document Detail


Activation of specific apoptotic caspases with an engineered small-molecule-activated protease.
MedLine Citation:
PMID:  20723762     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Apoptosis is a conserved cellular pathway that results in the activation of cysteine-aspartyl proteases, or caspases. To dissect the nonredundant roles of the executioner caspase-3, -6, and -7 in orchestrating apoptosis, we have developed an orthogonal protease to selectively activate each isoform in human cells. Our approach uses a split-tobacco etch virus (TEV) protease under small-molecule control, which we call the SNIPer, with caspase alleles containing genetically encoded TEV cleavage sites. These studies reveal that all three caspases are transiently activated but only activation of caspase-3 or -7 is sufficient to induce apoptosis. Proteomic analysis shown here and from others reveals that 20 of the 33 subunits of the 26S proteasome can be cut by caspases, and we demonstrate synergy between proteasome inhibition and dose-dependent caspase activation. We propose a model of proteolytic reciprocal negative regulation with mechanistic implications for the combined clinical use of proteasome inhibitors and proapoptotic drugs.
Authors:
Daniel C Gray; Sami Mahrus; James A Wells
Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Cell     Volume:  142     ISSN:  1097-4172     ISO Abbreviation:  Cell     Publication Date:  2010 Aug 
Date Detail:
Created Date:  2010-08-20     Completed Date:  2010-09-14     Revised Date:  2011-02-15    
Medline Journal Info:
Nlm Unique ID:  0413066     Medline TA:  Cell     Country:  United States    
Other Details:
Languages:  eng     Pagination:  637-46     Citation Subset:  IM    
Copyright Information:
Copyright 2010 Elsevier Inc. All rights reserved.
Affiliation:
Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, CA 94158, USA.
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MeSH Terms
Descriptor/Qualifier:
Apoptosis*
Caspase 3 / metabolism*
Caspase 6 / metabolism
Caspase 7 / metabolism*
Cell Line
Drug Design
Endopeptidases / genetics*,  metabolism*
Enzyme Activation / drug effects
Humans
Isoenzymes / metabolism
Leupeptins / pharmacology
Proteasome Endopeptidase Complex / antagonists & inhibitors
Protein Engineering*
Grant Support
ID/Acronym/Agency:
CA136779/CA/NCI NIH HHS; R01 CA136779-03/CA/NCI NIH HHS; R01 GM081051/GM/NIGMS NIH HHS; R01 GM081051-05/GM/NIGMS NIH HHS
Chemical
Reg. No./Substance:
0/Isoenzymes; 0/Leupeptins; 133407-82-6/benzyloxycarbonylleucyl-leucyl-leucine aldehyde; EC 3.4.-/Endopeptidases; EC 3.4.-/TEV protease; EC 3.4.22.-/Caspase 3; EC 3.4.22.-/Caspase 6; EC 3.4.22.-/Caspase 7; EC 3.4.25.1/Proteasome Endopeptidase Complex; EC 3.4.99.-/ATP dependent 26S protease
Comments/Corrections
Comment In:
Nat Methods. 2010 Oct;7(10):786   [PMID:  20936776 ]

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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