Open AccessExpression, purification, crystallization and preliminary X‐ray characterization of a putative glycosyltransferase of the GT‐A fold found in mycobacteria
Author(s) -
Fulton Zara,
Crellin Paul K.,
Brammananth Rajini,
ZakerTabrizi Leyla,
Coppel Ross L.,
Rossjohn Jamie,
Beddoe Travis
Publication year - 2008
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309108011196
Subject(s) - glycosyltransferase , biosynthesis , crystallization , enzyme , biochemistry , glycosidic bond , cloning (programming) , uridine diphosphate , chemistry , biology , organic chemistry , computer science , programming language
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 Å resolution using synchrotron radiation from a crystal belonging to space group P 4 1 2 1 2.