N-Ac-DEVD-N‘-(Polyfluorobenzoyl)−R110:  Novel Cell-Permeable Fluorogenic Caspase Substrates for the Detection of Caspase Activity and Apoptosis

• Published in: Bioconjugate chemistry Vol. 14; no. 2; pp. 458 - 463
• Main Authors: Zhang, Han-Zhong, Kasibhatla, Shailaja, Guastella, John, Tseng, Ben, Drewe, John, Cai, Sui Xiong
• Format: Journal Article
• Language: English
• Published: United States American Chemical Society 01.03.2003
• Subjects: Apoptosis - drug effects; Carcinogens - pharmacology; Caspase 3; Caspases - chemistry; Caspases - metabolism; Cell Membrane Permeability; Coumarins - chemistry; Flow Cytometry; Fluorescent Dyes - chemistry; HL-60 Cells; Humans; Magnetic Resonance Spectroscopy; Oligopeptides - chemistry; Recombinant Proteins - chemistry; Rhodamines - chemistry; Staurosporine - pharmacology; Substrate Specificity
• ISSN: 1043-1802; 1520-4812
• DOI: 10.1021/bc0256188

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.