Characterization of Arabinosyl Transfer Reactions in the Biosynthesis of Mycobacterial Cell Envelope (Lipo)Polysaccharides

Methods Mol Biol. 2019:1954:175-186. doi: 10.1007/978-1-4939-9154-9_14.

Abstract

D-Arabinofuranose is a major glycosyl constituent of mycobacteria found in two essential cell envelope heteropolysaccharides, arabinogalactan and lipoarabinomannan. Seven different arabinosyltransferases at least are required to synthesize the arabinan domain of these two major glycans. Because of their interest from the perspective of drug development, these enzymes have been the object of intense investigations. In this chapter, we describe the protocols used to perform nonradioactive arabinosyltransferase assays with synthetic acceptor and donor substrates and characterize the enzymatic products of the reactions by mass spectrometry.

Keywords: Arabinosyltransferase; D-Arabinose; Lipid donor; Mycobacteria; Synthetic arabinoside acceptor.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Arabinose / analogs & derivatives
  • Arabinose / metabolism
  • Bacterial Proteins / metabolism*
  • Biosynthetic Pathways
  • Cell Wall / enzymology
  • Cell Wall / metabolism
  • Chromatography, Liquid / methods
  • Enzyme Assays / methods*
  • Galactans / metabolism*
  • Lipopolysaccharides / metabolism*
  • Mass Spectrometry / methods
  • Mycobacterium smegmatis / enzymology
  • Mycobacterium smegmatis / metabolism*
  • Pentosyltransferases / metabolism*
  • Substrate Specificity

Substances

  • Bacterial Proteins
  • Galactans
  • Lipopolysaccharides
  • arabinofuranose
  • lipoarabinomannan
  • Arabinose
  • Pentosyltransferases
  • arabinosyltransferase
  • arabinogalactan