ORGN 536 |
| Zhi-Qiang Wang1, Jinfang Liao1, and Zhenjun Diwu2. (1) Reagent and Assay R&D, Molecular Devices Corp, 1311 Orleans Drive, Sunnyvale, CA 94089, (2) Reagent and Assay R&D, Molecular Devices Corporation, 1311 Orleans Drive, Sunnyvale, CA 94089 |
| Detection technologies for proteases are essential in the discovery of drug candidates for treatment and management of protease-mediated diseases. Fluorometric methods have been increasingly used for assaying various proteases, particularly caspases, which are important markers of cell death or apoptosis and potential targets for cancers and neurodegenerative diseases. Most of existing fluorogenic protease substrates lack the required sensitivity, among them, Rh110-based substrates are considered to be the most sensitive. Unfortunately Rh110 substrates generally have complex kinetics due to the two-step enzymatic reactions. Here, we report the synthesis and application of novel fluorometric substrates “DEVD-Rh110-MLP” for Caspase-3/7. Rhodamine-110 derivatives of caspase 3/7 substrates such as (Z-DEVD)2Rh110 have been previously used in cell-based apoptosis assays for detection of caspase 3/7. Our results indicated that DEVD-Rh110-MLP was converted or processed more efficiently in a single-step cleavage by Caspase-3 than (Z-DEVD)2Rh110. The improved dynamic change in fluorescence signal generated through cleavage of Z-DEVD-Rh110-MLP by Caspase-3/7 makes it an excellent tool for high throughput screening of compound libraries for specific Caspase-3/7 modulator(s).
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Bioorganic, Molecular Recognition, Asymmetric Reactions and Syntheses
11:00 AM-1:00 PM, Wednesday, September 10, 2003 Javits Convention Center -- Hall 1B/1C, Poster
Division of Organic Chemistry |