Chemical proteomics affinity chromatography

An increasing number of available protein-binding assays, functional cell-based assays, and methods of chemical proteomics (affinity chromatography, three-hybrid assays, pull-down assays) will allow for a better assignment of the specificity and selectivity of a hit compound. It would be desirable that the data collected during these screening... 

Katayama, H., Oda, Y. (2007). Chemical proteomics for drug discovery based on compound-immobilized affinity chromatography. J. Chromatogr. B, 855, 21-27. 

Chemical proteomics consists of the classical drug-affinity chromatography and modern high-resolution MS analysis for protein identification [3, 11]. The procedure typically involves immobilization of the compound of interest to a solid support through a spacer arm, and the affinity matrix is then used to purify specific interacting proteins from cellular lysate. The complex proteomic mixture is then proteolytically digested, and the resulting peptides are sequenced... 

Besides protein microarray proteome profiling, the term chemical proteomics (also chemoproteomics or pull-downs ) is mostly used in reference to the application of affinity chromatography protein purification when small molecules are the bait, and liquid chromatography separation of peptides precludes mass spectrometry as the universal readout (LC-MS/MS). 

Fig. 1 SILAC-based affinity chromatography for identification of the target proteins of small molecules, (a) Generation of two cell populations differentially labeled with light or heavy amino acids, (b) Employment of the metabolically labeled cell lysates from (a) in affinity-based chemical proteomics...

Breccia JD, Moran AC, Castro GR et al. (1998) Thermal stabiUzation by polyols of P-xyltmase from Bacillus amyloliquefaciens. J Chem Technol Biotechnol 71(3) 241-245 Breddam K, Beenfeldt T (1991) Acceleration of yeast autolysis by chemical methods for production of intracellular enzjnnes. J Appl Microbiol Biotechnol 35(3) 323-329 Brocklehurst K, Courey AJ, Gul S et al. (2004) Affinity and immunoaffinity chromatography. In Simpson RJ (ed). Purifying proteins for proteomics a laboratory manual CSHL Press, Cold Spring Harbor, pp 221-274... 

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