Sulfatase-activated fluorophores for rapid discrimination of mycobacterial species and strains

Proc Natl Acad Sci U S A. 2013 Aug 6;110(32):12911-6. doi: 10.1073/pnas.1222041110. Epub 2013 Jul 22.

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.

Keywords: Mycobacterium tuberculosis; chemical biology; enzyme assay; fluorescence; hydrolase.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acridones / chemistry*
  • Animals
  • Bacterial Proteins / metabolism
  • CHO Cells
  • COS Cells
  • Cell Line
  • Cell Line, Tumor
  • Chlorocebus aethiops
  • Cricetinae
  • Cricetulus
  • Electrophoresis / methods
  • Fluorescent Dyes / chemistry*
  • Gels
  • HEK293 Cells
  • Humans
  • Jurkat Cells
  • Molecular Structure
  • Mycobacterium / classification
  • Mycobacterium / enzymology
  • Mycobacterium / metabolism*
  • Mycobacterium tuberculosis / enzymology
  • Mycobacterium tuberculosis / metabolism
  • Species Specificity
  • Substrate Specificity
  • Sulfatases / metabolism*
  • Sulfates / chemistry
  • Sulfates / metabolism*
  • Sulfuric Acid Esters / chemistry*
  • Tuberculosis / diagnosis
  • Tuberculosis / microbiology

Substances

  • 7-hydroxy-9H-(1,3-dichloro-9,9-dimethylacridin-2-one)sulfate
  • Acridones
  • Bacterial Proteins
  • Fluorescent Dyes
  • Gels
  • Sulfates
  • Sulfuric Acid Esters
  • Sulfatases