| Purification of catalytically active caspase-12 and its biochemical characterization. | |
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MedLine Citation:
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PMID: 20646990 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Caspase-12, mainly detected in endoplasmic reticulum (ER), has been suggested to play a role in ER-mediated apoptosis and inflammatory caspase activation pathway. Cleavage of the prodomain by caspase-3/-7 at the carboxyl terminus of Asp94 or m-calpain at the carboxyl terminus of Lys158 was reported to be a part of caspase-12-involved apoptosis. We biochemically characterized the prodomain-free forms of caspase-12 and the equivalent enzymes; Deltapro1(G95-D419), rev-Deltapro1[(T319-N419)-(G95-D318), a reverse form of Deltapro1] and rev-Deltapro2[(T319-N419)-(T159-D318)]. The three variants showed comparable activities which were dependent on salt concentration and pH. Auto-proteolytic cleavage was observed at two sites (carboxyl termini of Asp318 and Asp320) in Deltapro1. Constitutively active forms of caspase-12 (rev-Deltapro1 and rev-Deltapro2) could induce cell death in cells transfected with the corresponding expression vectors, but no cleavage of caspase-3, DFF45 or Bid was observed, indicating caspase-12 may mediate a distinct apoptotic pathway rather than caspase-8 or -9-mediated cell death. |
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Authors:
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Hyun-Jung Lee; Sung Haeng Lee; Sung-Hee Park; M Golam Sharoar; Song Yub Shin; Jung Sup Lee; Byungyun Cho; Il-Seon Park |
Publication Detail:
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Type: In Vitro; Journal Article; Research Support, Non-U.S. Gov't Date: 2010-07-18 |
Journal Detail:
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Title: Archives of biochemistry and biophysics Volume: 502 ISSN: 1096-0384 ISO Abbreviation: Arch. Biochem. Biophys. Publication Date: 2010 Oct |
Date Detail:
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Created Date: 2010-08-31 Completed Date: 2010-09-20 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 0372430 Medline TA: Arch Biochem Biophys Country: United States |
Other Details:
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Languages: eng Pagination: 68-73 Citation Subset: IM |
Copyright Information:
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Copyright 2010 Elsevier Inc. All rights reserved. |
Affiliation:
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Department of Biology, Ewha Womans University, Seoul, Republic of Korea. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Apoptosis
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physiology Base Sequence Binding Sites Caspase 12 / chemistry, genetics, isolation & purification*, metabolism Caspase 7 / genetics, isolation & purification, metabolism Cell Line DNA Primers / genetics Genetic Variation Humans Hydrogen-Ion Concentration Osmolar Concentration Protein Structure, Tertiary Recombinant Proteins / genetics, isolation & purification, metabolism Transfection |
| Chemical | |
Reg. No./Substance:
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0/CASP12 protein, human; 0/DNA Primers; 0/Recombinant Proteins; EC 3.4.22.-/CASP7 protein, human; EC 3.4.22.-/Caspase 12; EC 3.4.22.-/Caspase 7 |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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