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Caspase-specific and nonspecific in vivo protein processing during Fas-induced apoptosis

Petra Van Damme (UGent) , Lennart Martens (UGent) , Jozef Van Damme (UGent) , Koen Hugelier, An Staes (UGent) , Joël Vandekerckhove (UGent) and Kris Gevaert (UGent)
(2005) NATURE METHODS. 2(10). p.771-777
Author
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Abstract
We generated a comprehensive picture of protease substrates in anti-Fas-treated apoptotic human Jurkat T lymphocytes. We used combined fractional diagonal chromatography (COFRADIC) sorting of protein amino-terminal peptides coupled to oxygen-16 or oxygen-18 differential labeling. We identified protease substrates and located the exact cleavage sites within processed proteins. Our analysis yielded 1,834 protein identifications and located 93 cleavage sites in 71 proteins. Indirect evidence of apoptosis-specific cleavage within 21 additional proteins increased the total number of processed proteins to 92. Most cleavages were at caspase consensus sites; however, other cleavage specificities suggest activation of other proteases. We validated several new processing events by immunodetection and by an in vitro assay using recombinant caspases and synthetic peptides containing presumed cleavage sites. The spliceosome complex appeared a preferred target, as 14 of its members were processed. Differential isotopic Labeling further revealed specific release of nucleosomal components from apoptotic nuclei.
Keywords
CLEAVAGE, CHROMATIN CONDENSATION, INHIBITION, PROTEASES, PEPTIDES, DISEASE, TARGETS, CLONING, MICE DEFICIENT, IDENTIFICATION

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Chicago
Van Damme, Petra, Lennart Martens, Jozef Van Damme, Koen Hugelier, An Staes, Joël Vandekerckhove, and Kris Gevaert. 2005. “Caspase-specific and Nonspecific in Vivo Protein Processing During Fas-induced Apoptosis.” Nature Methods 2 (10): 771–777.
APA
Van Damme, Petra, Martens, L., Van Damme, J., Hugelier, K., Staes, A., Vandekerckhove, J., & Gevaert, K. (2005). Caspase-specific and nonspecific in vivo protein processing during Fas-induced apoptosis. NATURE METHODS, 2(10), 771–777.
Vancouver
1.
Van Damme P, Martens L, Van Damme J, Hugelier K, Staes A, Vandekerckhove J, et al. Caspase-specific and nonspecific in vivo protein processing during Fas-induced apoptosis. NATURE METHODS. 2005;2(10):771–7.
MLA
Van Damme, Petra, Lennart Martens, Jozef Van Damme, et al. “Caspase-specific and Nonspecific in Vivo Protein Processing During Fas-induced Apoptosis.” NATURE METHODS 2.10 (2005): 771–777. Print.
@article{339248,
  abstract     = {We generated a comprehensive picture of protease substrates in anti-Fas-treated apoptotic human Jurkat T lymphocytes. We used combined fractional diagonal chromatography (COFRADIC) sorting of protein amino-terminal peptides coupled to oxygen-16 or oxygen-18 differential labeling. We identified protease substrates and located the exact cleavage sites within processed proteins. Our analysis yielded 1,834 protein identifications and located 93 cleavage sites in 71 proteins. Indirect evidence of apoptosis-specific cleavage within 21 additional proteins increased the total number of processed proteins to 92. Most cleavages were at caspase consensus sites; however, other cleavage specificities suggest activation of other proteases. We validated several new processing events by immunodetection and by an in vitro assay using recombinant caspases and synthetic peptides containing presumed cleavage sites. The spliceosome complex appeared a preferred target, as 14 of its members were processed. Differential isotopic Labeling further revealed specific release of nucleosomal components from apoptotic nuclei.},
  author       = {Van Damme, Petra and Martens, Lennart and Van Damme, Jozef and Hugelier, Koen and Staes, An and Vandekerckhove, Joël and Gevaert, Kris},
  issn         = {1548-7091},
  journal      = {NATURE METHODS},
  keywords     = {CLEAVAGE,CHROMATIN CONDENSATION,INHIBITION,PROTEASES,PEPTIDES,DISEASE,TARGETS,CLONING,MICE DEFICIENT,IDENTIFICATION},
  language     = {eng},
  number       = {10},
  pages        = {771--777},
  title        = {Caspase-specific and nonspecific in vivo protein processing during Fas-induced apoptosis},
  url          = {http://dx.doi.org/10.1038/NMETH792},
  volume       = {2},
  year         = {2005},
}

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