AI Summary
Chemical probes for bacterial glycosyltransferases are important for tracking expression levels and identifying strains. Existing probes based on sugar-nucleotides are limited in their use in intact cells. In this study, a new covalent probe for the bacterial galactosyltransferase LgtC was developed. The probe was able to label the enzyme in different forms and could potentially be used for developing LgtC-specific probes for bacterial strain detection. The stability of this probe in aqueous media and the unexpected improvement in activity are also discussed.
Chemical probes for bacterial glycosyltransferases are of interest for applications such as tracking of expression levels, and strain profiling and identification. Existing probes for glycosyltransferases are typically based on sugar-nucleotides, whose charged nature limits their applicability in intact cells. We report the development of an uncharged covalent probe for the bacterial galactosyltransferase LgtC, and its application for the fluorescent labelling of this enzyme in recombinant form, cell lysates, and intact cells. The probe was designed by equipping a previously reported covalent LgtC inhibitor based on a pyrazol-3-one scaffold with a 7-hydroxycoumarin fluorophore. We show that this pyrazol-3-ones scaffold is surprisingly stable in aqueous media, which may have wider implications for the use of pyrazol-3-ones as chemical probes. We also show that the 7-hydroxycoumarin fluorophore leads to an unexpected improvement in activity, which could be exploited for the development of second generation analogues. These results will provide a basis for the development of LgtC-specific probes for the detection of LgtC-expressing bacterial strains.
This article is Open Access
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