Real-Time Monitoring of Apoptosis by Caspase-3-Like Protease Induced FRET Reduction Triggered by Amyloid Aggregation
Joint Authors
Köhler, Martin
Berggren, Per-Olof
Schultz, Sebastian W.
Leibiger, Ingo
Paulsson, Johan F.
Westermark, Gunilla T.
Source
Issue
Vol. 2008, Issue 2008 (31 Dec. 2008), pp.1-12, 12 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2008-05-06
Country of Publication
Egypt
No. of Pages
12
Main Subjects
Abstract EN
Amyloid formation is cytotoxic and can activate the caspase cascade.
Here, we monitor caspase-3-like activity as reduction of fluorescence resonance energy transfer (FRET) using the contstruct pFRET2-DEVD containing enhanced cyan fluorescent protin (EYFP) linked by the caspase-3 specific cleavage site residues DEVD.
Beta-TC-6 cells were transfected, and the fluoorescence was measured at 440 nm excitation and 535 nm (EYFP) and 480 nm (ECFP) emission wavelength.
Cells were incubated with recombinant pro lset Amyloid Polypeptide (rec prolAPP) or the processing metabolites of prolAPP; the N-terminal flanking peptide withIAPP (recN+IAPP); IAPP with the C-terminal flanking peptied (recIAPP+C) and lslet Amyloid Polypeptide (recIAPP) .
Peptides were added in solubilized from (50 μM) or as performed amyloid-like fibrils, or as a combination of these.
FRET was measured and incubation with a mixture of solubilized peptide and performed fibrils resulted in loss of FRET and apoptosis was determined to occure in cells incubated with recproIAPP (49%), recN+IAPP (46%), recIAPP (72%) and recIAPP+C (59%).
These results show that proIAPP and the processing intermediates reside the same cell toxic capacity as IAPP, and they can all have a central role in the reduction of beta-cell number in type 2 diabetes.
American Psychological Association (APA)
Paulsson, Johan F.& Schultz, Sebastian W.& Köhler, Martin& Leibiger, Ingo& Berggren, Per-Olof& Westermark, Gunilla T.. 2008. Real-Time Monitoring of Apoptosis by Caspase-3-Like Protease Induced FRET Reduction Triggered by Amyloid Aggregation. Journal of Diabetes Research،Vol. 2008, no. 2008, pp.1-12.
https://search.emarefa.net/detail/BIM-988025
Modern Language Association (MLA)
Paulsson, Johan F.…[et al.]. Real-Time Monitoring of Apoptosis by Caspase-3-Like Protease Induced FRET Reduction Triggered by Amyloid Aggregation. Journal of Diabetes Research No. 2008 (2008), pp.1-12.
https://search.emarefa.net/detail/BIM-988025
American Medical Association (AMA)
Paulsson, Johan F.& Schultz, Sebastian W.& Köhler, Martin& Leibiger, Ingo& Berggren, Per-Olof& Westermark, Gunilla T.. Real-Time Monitoring of Apoptosis by Caspase-3-Like Protease Induced FRET Reduction Triggered by Amyloid Aggregation. Journal of Diabetes Research. 2008. Vol. 2008, no. 2008, pp.1-12.
https://search.emarefa.net/detail/BIM-988025
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-988025