Automated protein-ligand interaction screening by mass spectrometry
| Autor: | John Robert Porter, Richard J. K. Taylor, John Crosby, Matthew P. Crump, Rachel A. Garlish, Hannah J. Maple, Christine E. Prosser, Alistair James Henry, Jeff Kennedy, Ian Whitcombe, Laura Rigau-Roca |
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| Rok vydání: | 2011 |
| Předmět: |
Models
Molecular Quantitative structure–activity relationship Spectrometry Mass Electrospray Ionization Magnetic Resonance Spectroscopy bcl-X Protein Quantitative Structure-Activity Relationship Calorimetry Mass spectrometry Ligands Drug Discovery High-Throughput Screening Assays Wide dynamic range Animals Nanotechnology Throughput (business) Chromatography Drug discovery Chemistry Dynamic range Molecular Medicine Pyrazoles Muramidase Biological system Chickens Protein ligand |
| Zdroj: | Journal of medicinal chemistry. 55(2) |
| ISSN: | 1520-4804 |
| Popis: | Identifying protein-ligand binding interactions is a key step during early-stage drug discovery. Existing screening techniques are often associated with drawbacks such as low throughput, high sample consumption, and dynamic range limitations. The increasing use of fragment-based drug discovery (FBDD) demands that these techniques also detect very weak interactions (mM K(D) values). This paper presents the development and validation of a fully automated screen by mass spectrometry, capable of detecting fragment binding into the millimolar K(D) range. Low sample consumption, high throughput, and wide dynamic range make this a highly attractive, orthogonal approach. The method was applied to screen 157 compounds in 6 h against the anti-apoptotic protein target Bcl-x(L). Mass spectrometry results were validated using STD-NMR, HSQC-NMR, and ITC experiments. Agreement between techniques suggests that mass spectrometry offers a powerful, complementary approach for screening. |
| Databáze: | OpenAIRE |
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