N-Ac-DEVD-N'-(Polyfluorobenzoyl)-R110: novel cell-permeable fluorogenic caspase substrates for the detection of caspase activity and apoptosis.
N-Pentafluorobenzoyl-R110 (1a) and N-(2,3,4,5-tetrafluorobenzoyl)-R110 (1b) with enhanced cell retention properties, were prepared from rhodamine 110 (R-110) and the corresponding polyfluorobenzoyl chloride. N-Ac-DEVD-N'-pentafluorobenzoyl-R110 (3a) and N-Ac-DEVD-N'-(2,3,4,5-tetrafluorobenzoyl)-R110 (3b) were prepared as tetrapeptide substrates for caspases. Substrate 3b was efficiently cleaved by human recombinant caspase-3 in an enzyme assay. Substrate 3b also was efficiently cleaved in cell-based apoptosis assays. After cleavage in apoptotic cells by activated caspases, the substrate becomes fluorescent as measured by flow cytometry. These substrates should prove useful in cell-based assays for studying apoptosis inducers and inhibitors.
Zhang
Han-Zhong
HZ
Maxim Pharmaceuticals, 6650 Nancy Ridge Drive, San Diego, California 92121, USA.
Kasibhatla
Shailaja
S
Guastella
John
J
Tseng
Ben
B
Drewe
John
J
Cai
Sui Xiong
SX
D017209:Apoptosis / Q000187:drug effects*; D002273:Carcinogens / Q000494:pharmacology; D053148:Caspase 3; D020169:Caspases / Q000737:chemistry / Q000378:metabolism*; D002463:Cell Membrane Permeability; D003374:Coumarins / Q000737:chemistry*; D005434:Flow Cytometry; D005456:Fluorescent Dyes / Q000737:chemistry*; D018922:HL-60 Cells; D006801:Humans; D009682:Magnetic Resonance Spectroscopy; D009842:Oligopeptides / Q000737:chemistry*; D011994:Recombinant Proteins / Q000737:chemistry; D012235:Rhodamines / Q000737:chemistry; D019311:Staurosporine / Q000494:pharmacology; D013379:Substrate Specificity
D002273:Carcinogens; D003374:Coumarins; D005456:Fluorescent Dyes; C478643:N-Ac-DEVD-N'-(Polyfluorobenzoyl)-R110; D009842:Oligopeptides; D011994:Recombinant Proteins; D012235:Rhodamines; C505407:CASP3 protein, human; D053148:Caspase 3; D020169:Caspases; D019311:Staurosporine