RESUMO
A strategy for labeling native enzymes in a manner that preserves their activity is reported: capture-tag-release (CTR). Key to this approach is the small molecule CTR probe that contains an enzyme inhibitor, benzophenone crosslinker, and aryl phosphine ester. After UV-derived capture of the enzyme, addition of an azide-containing tag triggers a Staudinger ligation that labels the enzyme. A further consequence of the Staudinger ligation is fragmentation of the CTR probe, thus releasing the inhibitor and restoring enzymatic activity. As a proof-of-principle, the CTR strategy was applied to the hydrolase ß-galactosidase. The enzyme was efficiently labeled with biotin, and the kinetic data for the biotinylated enzyme were comparable to those for unlabeled ß-galactosidase. The CTR probe exhibits excellent targeting specificity, as it selectively labeled ß-galactosidase in a complex protein mixture.
