title: Sulfatase-activated fluorophores for rapid discrimination of mycobacterial species and strains.
abstract: Most current diagnostic tests for tuberculosis do not reveal the species or strain of pathogen causing pulmonary infection, which can lead to inappropriate treatment regimens and the spread of disease. Here, we report an assay for mycobacterial strain assignment based on genetically conserved mycobacterial sulfatases. We developed a sulfatase-activated probe, 7-hydroxy-9H-(1,3-dichloro-9,9-dimethylacridin-2-one)-sulfate, that detects enzyme activity in native protein gels, allowing the rapid detection of sulfatases in mycobacterial lysates. This assay revealed that mycobacterial strains have distinct sulfatase fingerprints that can be used to judge both the species and lineage. Our results demonstrate the potential of enzyme-activated probes for rapid pathogen discrimination for infectious diseases.
authors.affiliation: Department of Chemistry, University of California, Berkeley, CA 94720-1460, USA.
authors.forename: Kimberly E
authors.initials: KE
authors.lastname: Beatty
authors.forename: Monique
authors.initials: M
authors.lastname: Williams
authors.forename: Brian L
authors.initials: BL
authors.lastname: Carlson
authors.forename: Benjamin M
authors.initials: BM
authors.lastname: Swarts
authors.forename: Robin M
authors.initials: RM
authors.lastname: Warren
authors.forename: Paul D
authors.initials: PD
authors.lastname: van Helden
authors.forename: Carolyn R
authors.initials: CR
authors.lastname: Bertozzi
mesh_terms: D054831:Acridones / Q000737:chemistry*; D000818:Animals; D001426:Bacterial Proteins / Q000378:metabolism; D016466:CHO Cells; D019556:COS Cells; D002460:Cell Line; D045744:Cell Line, Tumor; D002522:Chlorocebus aethiops; D006224:Cricetinae; D003412:Cricetulus; D004586:Electrophoresis / Q000379:methods; D005456:Fluorescent Dyes / Q000737:chemistry*; D005782:Gels; D057809:HEK293 Cells; D006801:Humans; D019169:Jurkat Cells; D015394:Molecular Structure; D009161:Mycobacterium / Q000145:classification / Q000201:enzymology / Q000378:metabolism*; D009169:Mycobacterium tuberculosis / Q000201:enzymology / Q000378:metabolism; D013045:Species Specificity; D013379:Substrate Specificity; D013429:Sulfatases / Q000378:metabolism*; D013431:Sulfates / Q000737:chemistry / Q000378:metabolism*; D013463:Sulfuric Acid Esters / Q000737:chemistry*; D014376:Tuberculosis / Q000175:diagnosis / Q000382:microbiology
chemical_list: C584059:7-hydroxy-9H-(1,3-dichloro-9,9-dimethylacridin-2-one)sulfate; D054831:Acridones; D001426:Bacterial Proteins; D005456:Fluorescent Dyes; D005782:Gels; D013431:Sulfates; D013463:Sulfuric Acid Esters; D013429:Sulfatases
keywords: Mycobacterium tuberculosis; chemical biology; enzyme assay; fluorescence; hydrolase