Biochemical properties of Neisseria gonorrhoeae LgtE.
A fragment of chromosomal DNA encoding the lgtE gene of Neisseria gonorrhoeae strain F62 was amplified by PCR and cloned into the expression vector pET15b. Functional LgtE was purified and its biochemical properties were determined. The purified enzyme was maximally active in buffer containing manganese; minimal activity was obtained in buffer containing other divalent cations. LgtE was only able to mediate the addition of UDP-galactose into neisserial lipooligosaccharides (LOSs). We used a variety of genetically defined and chemically verified LOS structures to determine acceptor specificity. LgtE was able to mediate the addition of galactose into a variety of LOS structures, indicating the this enzyme possesses broad acceptor specificity. Furthermore, it was able to add multiple galactose residues onto LOS. We also determined that this enzyme was capable of adding galactose onto both the alpha and beta chains of neisserial LOS.
Piekarowicz
Andrzej
A
Institute of Microbiology, University of Warsaw, 02-096 Warsaw, Poland.
Stein
Daniel C
DC
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D002240:Carbohydrate Sequence; D003470:Culture Media; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D005690:Galactose / Q000378:metabolism; D005700:Galactosyltransferases / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D016695:Glycosyltransferases / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D008070:Lipopolysaccharides / Q000737:chemistry / Q000378:metabolism; D008969:Molecular Sequence Data; D009344:Neisseria gonorrhoeae / Q000201:enzymology* / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D013379:Substrate Specificity
D001426:Bacterial Proteins; D003470:Culture Media; D008070:Lipopolysaccharides; D011994:Recombinant Proteins; C023023:lipid-linked oligosaccharides; D016695:Glycosyltransferases; D005700:Galactosyltransferases; C469906:LgtE protein, Neisseria gonorrhoeae; D005690:Galactose