Expression, purification, crystallization and preliminary X-ray characterization of a putative glycosyltransferase of the GT-A fold found in mycobacteria Academic Article uri icon

abstract

  • Glycosidic bond formation is a ubiquitous enzyme-catalysed reaction. This glycosyltransferase-mediated process is responsible for the biosynthesis of innumerable oligosaccharides and glycoconjugates and is often organism- or cell-specific. However, despite the abundance of genomic information on glycosyltransferases (GTs), there is a lack of structural data for this versatile class of enzymes. Here, the cloning, expression, purification and crystallization of an essential 329-amino-acid (34.8 kDa) putative GT of the classic GT-A fold implicated in mycobacterial cell-wall biosynthesis are reported. Crystals of MAP2569c from Mycobacterium avium subsp. paratuberculosis were grown in 1.6 M monoammonium dihydrogen phosphate and 0.1 M sodium citrate pH 5.5. A complete data set was collected to 1.8 A resolution using synchrotron radiation from a crystal belonging to space group P4(1)2(1)2.

authors

  • Fulton, Zara
  • Crellin, Paul K
  • Brammananth, Rajini
  • Zaker-Tabrizi, Leyla
  • Coppel, Ross L
  • Rossjohn, Jamie
  • Beddoe, Travis

publication date

  • May 1, 2008