An expanded set of fluorogenic sulfatase activity probes

Fluorogenic probes that are activated by an enzymatic transformation are ideally suited for profiling enzyme activities in biological systems. Here, we describe two fluorogenic enzyme probes, 3-O-methylfluorescein-sulfate and resorufin-sulfate, that can be used to detect sulfatases in mycobacterial lysates. Both probes were validated with a set of commercial sulfatases and used to reveal species-specific sulfatase banding patterns in a gel-resolved assay of mycobacterial lysates. The fluorogenic probes described here are suitable for various assays and provide a starting point for creating new sulfatase probes with improved selectivity for mycobacterial sulfatases. Revealing sulfatases: Two new fluorogenic enzyme probes, 3-O-methylfluorescein sulfate and resorufin-sulfate, display a large increase in quantum yield upon hydrolysis by sulfatases. Both probes enable rapid, sensitive detection of sulfatase activity in a protein gel assay format, which allows the examination of sulfatases from a variety of mycobacterial pathogens.