Azido Pentoses: A New Tool To Efficiently Label Mycobacterium tuberculosis Clinical Isolates

Chembiochem. 2017 Jul 4;18(13):1172-1176. doi: 10.1002/cbic.201600706. Epub 2017 Mar 29.

Abstract

Mycobacterium tuberculosis (Mtb), the main causative agent of tuberculosis (Tb), has a complex cell envelope which forms an efficient barrier to antibiotics, thus contributing to the challenges of anti-tuberculosis therapy. However, the unique Mtb cell wall can be considered an advantage and be utilized to selectively label Mtb bacteria. Here we introduce three azido pentoses as new compounds for metabolic labeling of Mtb: 3-azido arabinose (3AraAz), 3-azido ribose (3RiboAz), and 5-azido arabinofuranose (5AraAz). 5AraAz demonstrated the highest level of Mtb labeling and was efficiently incorporated into the Mtb cell wall. All three azido pentoses can be easily used to label a variety of Mtb clinical isolates without influencing Mtb-dependent phagosomal maturation arrest in infection studies with human macrophages. Thus, this metabolic labeling method offers the opportunity to attach desired molecules to the surface of Mtb bacteria in order to facilitate investigation of the varying virulence characteristics of different Mtb clinical isolates, which influence the outcome of a Tb infection.

Keywords: Mycobacterium tuberculosis; arabinan metabolism; carbohydrates; click chemistry; metabolic labeling.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Azides / chemistry*
  • Biomarkers / metabolism
  • Cell Wall / chemistry*
  • Cell Wall / metabolism
  • Flow Cytometry
  • Gene Expression
  • Humans
  • Lysosomal Membrane Proteins / genetics
  • Lysosomal Membrane Proteins / immunology
  • Macrophages / cytology
  • Macrophages / immunology
  • Mycobacterium tuberculosis / chemistry*
  • Mycobacterium tuberculosis / metabolism
  • Pentoses / chemistry*
  • Phagocytosis
  • Staining and Labeling / methods*
  • rab5 GTP-Binding Proteins / genetics
  • rab5 GTP-Binding Proteins / immunology

Substances

  • Azides
  • Biomarkers
  • LAMP1 protein, human
  • Lysosomal Membrane Proteins
  • Pentoses
  • RAB5C protein, human
  • rab5 GTP-Binding Proteins