Abstract
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.
Original language | English (US) |
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Pages (from-to) | 1101-1105 |
Number of pages | 5 |
Journal | ChemBioChem |
Volume | 15 |
Issue number | 8 |
DOIs | |
State | Published - May 26 2014 |
Keywords
- Mycobacterium tuberculosis
- fluorescent probes
- hydrolases
- protein imaging
- sulfatases
ASJC Scopus subject areas
- Biochemistry
- Molecular Medicine
- Molecular Biology
- Organic Chemistry