By Gerard Drewes, Marcus Bantscheff
The multidisciplinary technology of chemical proteomics experiences how small molecules of man-made or typical foundation bind to proteins and modulate their functionality. In Chemical Proteomics: equipment and Protocols, specialist researchers within the box supply key concepts to enquire chemical proteomics concentrating on analytical ideas, how probes are generated, ideas for the invention of small molecule goals and the probing of goal functionality, and small molecule ligand and drug discovery. Written within the hugely profitable tools in Molecular Biology™ sequence structure, chapters contain introductions to their respective issues, lists of the required fabrics and reagents, step by step, conveniently reproducible laboratory protocols, and key pointers on troubleshooting and heading off identified pitfalls. Authoritative and sensible, Chemical Proteomics : equipment and Protocols seeks to supply methodologies that might give a contribution to a much broader software of chemical proteomics equipment in biochemical and cellphone organic laboratories.
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Extra resources for Chemical Proteomics: Methods and Protocols (Methods in Molecular Biology, v803)
2. Wash the gel two times with 30% ethanol for 20 min and for another 20 min with water (pro analysi). In the meantime prepare the sensitizer, the silver nitrate (refrigerator) and the developer. Fig. 3. SDS-PAGE-based analysis of the dasatinib pulldown eluate from KU812 CML cells (9). (a) Silver staining shows several bands that subsequent LC-MS/MS analysis reveals to correlate with validated dasatinib target kinases. (b) Immunoblot with anti-ABL 21–63 antibody shows the enrichment of BCR-ABL and c-ABL by the dasatinib affinity matrix while an ampicillin control matrix does not display affinity for the dasatinib cognate targets.
Chemical Proteomics: Methods and Protocols, Methods in Molecular Biology, vol. 1007/978-1-61779-364-6_3, © Springer Science+Business Media, LLC 2012 25 26 U. Rix et al. characterization of its molecular mechanism of action (1, 2). This approach requires the immobilization of the compound of interest on solid support as an early step. The strategy for immobilization has to be designed very carefully as chemical modification of any bioactive compound harbors the risk of impairing or abrogating its biological activity through disruption of the protein interaction interface.
23. 24. 25. 26. 27. target of thalidomide teratogenicity. Science. 327, 1345–1350. Fliri, A. , Loging, W. , Thadeio, P. , and Volkmann, R. A. (2005) Analysis of druginduced effect patterns to link structure and side effects of medicines. Nat. Chem. Biol. 1, 389–397. Ong, S. , Margolin, A. , Doud, M. , Mani, D. , Wood, J. , Tolliday, N. , Koehler, A. , Marcaurelle, L. , Golub, T. , Gould, R. , Schreiber, S. , and Carr, S. A. (2009) Identifying the proteins to which small-molecule probes and drugs bind in cells.