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Purification of catalytically active caspase-12 and its biochemical characterization

Authors
  • Lee, Hyun-Jung
  • Lee, Sung Haeng
  • Park, Sung-Hee
  • Sharoar, Md. Golam
  • Shin, Song Yub
  • Lee, Jung Sup
  • Cho, Byungyun
  • Park, Il-Seon
Type
Published Article
Journal
Archives of Biochemistry and Biophysics
Publisher
Elsevier BV
Publication Date
Jan 01, 2010
Volume
502
Issue
1
Pages
68–73
Identifiers
DOI: 10.1016/j.abb.2010.07.013
Source
Elsevier
Keywords
License
Unknown

Abstract

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; Δpro1(G95-D419), rev-Δpro1[(T319-N419)-(G95-D318), a reverse form of Δpro1] and rev-Δpro2[(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 Δpro1. Constitutively active forms of caspase-12 (rev-Δpro1 and rev-Δpro2) 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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