Imaging of caspase-3 activation in HeLa cells stimulated with etoposide using a novel fluorescent probe.
Microscopic visualization of intracellular enzyme activity can provide information about the physiological role of the enzyme. Caspases are cysteine proteases that have critical roles in the execution of apoptosis. General fluorometric substrates of caspase-3, such as DEVD-MCA, are unsuitable for imaging because they are excited at short wavelength, so we designed and synthesized novel fluorescent probes that are excited at suitable wavelengths for detecting caspase-3 activity in living cells. Using one of these probes, we succeeded in microscopic visualization of caspase-3-like activity within HeLa cells treated with etoposide. The caspase-3-like activity was increased in the cytosol at first, then expanded to the whole cell.
Graduate School of Pharmaceutical Sciences, The University of Tokyo, Japan.
S
S
Mizukami
K
K
Kikuchi
T
T
Higuchi
Y
Y
Urano
T
T
Mashima
T
T
Tsuruo
T
T
Nagano
D017209:Apoptosis; D053148:Caspase 3; D020169:Caspases / Q000302:isolation & purification* / Q000378:metabolism; D004789:Enzyme Activation; D005047:Etoposide / Q000494:pharmacology; D005456:Fluorescent Dyes; D006367:HeLa Cells; D006801:Humans; D008856:Microscopy, Fluorescence / Q000379:methods*; D059005:Topoisomerase II Inhibitors
D005456:Fluorescent Dyes; D059005:Topoisomerase II Inhibitors; D005047:Etoposide; C505407:CASP3 protein, human; D053148:Caspase 3; D020169:Caspases