title: Expression, purification, crystallization and preliminary X-ray characterization of a putative glycosyltransferase of the GT-A fold found in mycobacteria.
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.affiliation: The Protein Crystallography Unit, Department of Biochemistry and Molecular Biology, School of Biomedical Sciences, Monash University, Clayton, Victoria 3800, Australia.
authors.forename: Zara
authors.initials: Z
authors.lastname: Fulton
authors.forename: Paul K
authors.initials: PK
authors.lastname: Crellin
authors.forename: Rajini
authors.initials: R
authors.lastname: Brammananth
authors.forename: Leyla
authors.initials: L
authors.lastname: Zaker-Tabrizi
authors.forename: Ross L
authors.initials: RL
authors.lastname: Coppel
authors.forename: Jamie
authors.initials: J
authors.lastname: Rossjohn
authors.forename: Travis
authors.initials: T
authors.lastname: Beddoe
mesh_terms: D003001:Cloning, Molecular; D003460:Crystallization; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D016695:Glycosyltransferases / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D016927:Mycobacterium avium subsp. paratuberculosis / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development; D014961:X-Ray Diffraction
chemical_list: D016695:Glycosyltransferases