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Oxidized low-density lipoprotein induces calpain-dependent cell death and ubiquitination of caspase 3 in HMEC-1 endothelial cells.

Ares, Isabella LU ; Saido, Takaomi C; Andersson, Tommy LU and Ares, Mikko P S (2003) In Biochem J 374(Pt 2). p.403-411
Abstract
Oxidized low-density lipoprotein (oxLDL) is known to induce apoptosis in endothelial cells, and this is believed to contribute to the progression of atherosclerosis. In the present study we made the novel observation that oxLDL-induced death of HMEC-1 cells is accompanied by activation of calpain. The mu-calpain inhibitor PD 151746 decreased oxLDL-induced cytotoxicity, whereas the general caspase inhibitor BAF (t-butoxycarboryl-Asp-methoxyfluoromethylketone) had no effect. Also, oxLDL provoked calpain-dependent proteolysis of cytoskeletal a-fodrin in the HMEC-1 cells. Our observation of an autoproteolytic cleavage of the 80 kDa subunit of mu-calpain provided further evidence for an oxLDL-indunced stimulation of calpain activity. The Bcl-2... (More)
Oxidized low-density lipoprotein (oxLDL) is known to induce apoptosis in endothelial cells, and this is believed to contribute to the progression of atherosclerosis. In the present study we made the novel observation that oxLDL-induced death of HMEC-1 cells is accompanied by activation of calpain. The mu-calpain inhibitor PD 151746 decreased oxLDL-induced cytotoxicity, whereas the general caspase inhibitor BAF (t-butoxycarboryl-Asp-methoxyfluoromethylketone) had no effect. Also, oxLDL provoked calpain-dependent proteolysis of cytoskeletal a-fodrin in the HMEC-1 cells. Our observation of an autoproteolytic cleavage of the 80 kDa subunit of mu-calpain provided further evidence for an oxLDL-indunced stimulation of calpain activity. The Bcl-2 protein Bid was also cleaved during oxLDL-elicited cell death, and this was prevented by calpain inhibitors, but not by inhibitors of cathepsin B and caspases. Treating the HMEC-1 cells with oxLDL did not result in detectable activation of procaspase 3 or cleavage of PARP [poly(ADP-ribose) polymerase], but it did cause polyubiquitination of caspase 3, indicating inactivation and possible degradation of this protease. Despite the lack of caspase 3 activation, oxLDL treatment led to the formation of nucleosomal DNA fragments characteristic of apoptosis. These novel results show that oxLDL initiates a calpain-mediated death-signalling pathway in endothelial cells. (Less)
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organization
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Contribution to journal
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published
subject
keywords
Lipoproteins, Intracellular Fluid: metabolism, Hydrolysis, Human, Growth Inhibitors: toxicity, Enzyme Activation: drug effects, Vascular: cytology, Calcium: metabolism, Apoptosis: drug effects, Endothelium, Caspases: metabolism, Carrier Proteins: metabolism, Calpain: physiology, Cell Line, DNA Fragmentation: physiology, Vascular: physiology, Vascular: enzymology, Endopeptidases: physiology, LDL: toxicity, Microfilament Proteins: metabolism, Oxidation-Reduction, Proto-Oncogene Proteins c-bcl-2: metabolism, Support, Non-U.S. Gov't, Ubiquitin: metabolism
in
Biochem J
volume
374
issue
Pt 2
pages
403 - 411
publisher
Portland Press Limited
external identifiers
  • wos:000185317800013
  • pmid:12775216
  • scopus:0041829109
ISSN
1470-8728
DOI
10.1042/BJ20021955
language
English
LU publication?
yes
id
5dc0f5ce-362b-4dda-a4ba-ced603e0b8ad (old id 121783)
alternative location
http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=12775216&dopt=Abstract
date added to LUP
2007-07-26 15:43:57
date last changed
2018-01-07 09:25:54
@article{5dc0f5ce-362b-4dda-a4ba-ced603e0b8ad,
  abstract     = {Oxidized low-density lipoprotein (oxLDL) is known to induce apoptosis in endothelial cells, and this is believed to contribute to the progression of atherosclerosis. In the present study we made the novel observation that oxLDL-induced death of HMEC-1 cells is accompanied by activation of calpain. The mu-calpain inhibitor PD 151746 decreased oxLDL-induced cytotoxicity, whereas the general caspase inhibitor BAF (t-butoxycarboryl-Asp-methoxyfluoromethylketone) had no effect. Also, oxLDL provoked calpain-dependent proteolysis of cytoskeletal a-fodrin in the HMEC-1 cells. Our observation of an autoproteolytic cleavage of the 80 kDa subunit of mu-calpain provided further evidence for an oxLDL-indunced stimulation of calpain activity. The Bcl-2 protein Bid was also cleaved during oxLDL-elicited cell death, and this was prevented by calpain inhibitors, but not by inhibitors of cathepsin B and caspases. Treating the HMEC-1 cells with oxLDL did not result in detectable activation of procaspase 3 or cleavage of PARP [poly(ADP-ribose) polymerase], but it did cause polyubiquitination of caspase 3, indicating inactivation and possible degradation of this protease. Despite the lack of caspase 3 activation, oxLDL treatment led to the formation of nucleosomal DNA fragments characteristic of apoptosis. These novel results show that oxLDL initiates a calpain-mediated death-signalling pathway in endothelial cells.},
  author       = {Ares, Isabella and Saido, Takaomi C and Andersson, Tommy and Ares, Mikko P S},
  issn         = {1470-8728},
  keyword      = {Lipoproteins,Intracellular Fluid: metabolism,Hydrolysis,Human,Growth Inhibitors: toxicity,Enzyme Activation: drug effects,Vascular: cytology,Calcium: metabolism,Apoptosis: drug effects,Endothelium,Caspases: metabolism,Carrier Proteins: metabolism,Calpain: physiology,Cell Line,DNA Fragmentation: physiology,Vascular: physiology,Vascular: enzymology,Endopeptidases: physiology,LDL: toxicity,Microfilament Proteins: metabolism,Oxidation-Reduction,Proto-Oncogene Proteins c-bcl-2: metabolism,Support,Non-U.S. Gov't,Ubiquitin: metabolism},
  language     = {eng},
  number       = {Pt 2},
  pages        = {403--411},
  publisher    = {Portland Press Limited},
  series       = {Biochem J},
  title        = {Oxidized low-density lipoprotein induces calpain-dependent cell death and ubiquitination of caspase 3 in HMEC-1 endothelial cells.},
  url          = {http://dx.doi.org/10.1042/BJ20021955},
  volume       = {374},
  year         = {2003},
}