Rapid caspase-3 activation during apoptosis revealed using fluorescence-resonance energy transfer.

Abstract:

:Caspase-3 is a crucial component of the apoptotic machinery in many cell types. Here, we report the timescale of caspase-3 activation in single living cells undergoing apoptosis. This was achieved by measuring the extent of fluorescence resonance energy transfer within a recombinant substrate containing cyan fluorescent protein (CFP) linked by a short peptide possessing the caspase-3 cleavage sequence, DEVD, to yellow fluorescent protein (YFP; i.e. CFP-DEVD-YFP). We demonstrate that, once initiated, the activation of caspase-3 is a very rapid process, taking 5 min or less to reach completion. Furthermore, this process occurs almost simultaneously with a depolarization of the mitochondrial membrane potential. These events occur just prior to the characteristic morphological changes associated with apoptosis. Our results clearly demonstrate that, once initiated, the commitment of cells to apoptosis is a remarkably rapid event when visualized at the single cell level.

journal_name

EMBO Rep

journal_title

EMBO reports

authors

Tyas L,Brophy VA,Pope A,Rivett AJ,Tavaré JM

doi

10.1093/embo-reports/kvd050

keywords:

subject

Has Abstract

pub_date

2000-09-01 00:00:00

pages

266-70

issue

3

eissn

1469-221X

issn

1469-3178

journal_volume

1

pub_type

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