SITE-SPECIFIC LABELING OF E. coli CITRATE SYNTHASE WITH BROMOBIMANE

摘要

We were sucessful in finding conditions for putting an unique bimane label on the surface of a fully active CS protein. Because we were using folded protein, the variability in labeling under more extreme conditions can probably be explained by the protein structure. This aspect is under investigation. However, if one wished to label all the cyteines in a protein, it might be wise to denature it first. CHCA matrix was more tolerant of variation in laser fluence, but DHB would be better for a proteokics application where the ± 190 Da is very clear. Only ESI was gentle enough to retain the bimane label on the protein and its trypsin-produced peptides.