Biochemical analysis of Lgt3, a glycosyltransferase of the bacterium Moraxella catarrhalis.
The lipooligosaccharide (LOS) of Moraxella catarrhalis is unusual in that it lacks heptose. The sugar linking oligosaccharide to Lipid A is a trisubstituted glucose. A single enzyme, Lgt3, is suggested to trisubstitute this core sugar. The lgt3 gene encodes two distinct domains with high similarity to glucosyltransferases of the GT-A superfamily, thus encoding a bidomain, multifunctional glucosyltransferase. To characterise Lgt3, the gene was amplified from M. catarrhalis, expressed in Escherichia coli, and purified. Analysis of its glycosyltransferase catalytic activity ascertained the pH and temperature optima for Lgt3. The donor specificity and acceptor specificity were examined. This study confirms that Lgt3 is a glucosyltransferase which catalyses addition of glucose to its cognate receptor, a terminal glucose presented as the core region of LOS.
Institute for Glycomics, Griffith University, Gold Coast Campus, Southport, Qld 4222, Australia.
Isabelle
I
Faglin
Joe
J
Tiralongo
Jennifer C
JC
Wilson
Patrick M
PM
Collins
Ian R
IR
Peak
D002384:Catalysis; D004926:Escherichia coli / Q000235:genetics; D005947:Glucose / Q000378:metabolism; D016695:Glycosyltransferases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008050:Lipid A / Q000378:metabolism; D001936:Moraxella catarrhalis / Q000201:enzymology* / Q000235:genetics; D017434:Protein Structure, Tertiary; D013379:Substrate Specificity
D008050:Lipid A; D016695:Glycosyltransferases; D005947:Glucose