Oxidized low-density lipoprotein induces calpain-dependent cell death and ubiquitination of caspase 3 in HMEC-1 endothelial cells.
(2003) In Biochemical Journal 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)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/121783
- author
- Ares, Isabella LU ; Saido, Takaomi C ; Andersson, Tommy LU and Ares, Mikko P S
- organization
- publishing date
- 2003
- type
- Contribution to journal
- publication status
- 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
- Biochemical Journal
- volume
- 374
- issue
- Pt 2
- pages
- 403 - 411
- publisher
- Portland Press
- 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
- 2016-04-01 16:38:51
- date last changed
- 2022-04-22 23:28:58
@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}}, 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}}, language = {{eng}}, number = {{Pt 2}}, pages = {{403--411}}, publisher = {{Portland Press}}, series = {{Biochemical Journal}}, 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}}, doi = {{10.1042/BJ20021955}}, volume = {{374}}, year = {{2003}}, }