Multifunctionality of Campylobacter jejuni sialyltransferase CstII: characterization of GD3/GT3 oligosaccharide synthase, GD3 oligosaccharide sialidase, and trans-sialidase activities.
CstII from bacterium Campylobacter jejuni strain OH4384 has been previously characterized as a bifunctional sialyltransferase having both alpha2,3-sialyltransferase (GM3 oligosaccharide synthase) and alpha2,8-sialyltransferase (GD3 oligosaccharide synthase) activities which catalyze the transfer of N-acetylneuraminic acid (Neu5Ac) from cytidine 5'-monophosphate (CMP)-Neu5Ac to C-3' of the galactose in lactose and to C-8 of the Neu5Ac in 3'-sialyllactose, respectively (Gilbert M, Karwaski MF, Bernatchez S, Young NM, Taboada E, Michniewicz J, Cunningham AM, Wakarchuk WW. 2002. The genetic bases for the variation in the lipo-oligosaccharide of the mucosal pathogen, Campylobacter jejuni. Biosynthesis of sialylated ganglioside mimics in the core oligosaccharide. J Biol Chem. 277:327-337). We report here the characterization of a truncated CstII mutant (CstIIDelta32(I53S)) cloned from a synthetic gene whose codons are optimized for an Escherichia coli expression system. In addition to the alpha2,3- and alpha2,8-sialyltransferase activities reported before for the synthesis of GM3- and GD3-type oligosaccharides, respectively, the CstIIDelta32(I53S) has alpha2,8-sialyltransferase (GT3 oligosaccharide synthase) activity for the synthesis of GT3 oligosaccharide. It also has alpha2,8-sialidase (GD3 oligosaccharide sialidase) activity that catalyzes the specific cleavage of the alpha2,8-sialyl linkage of GD3-type oligosaccharides and alpha2,8-trans-sialidase (GD3 oligosaccharide trans-sialidase) activity that catalyzes the transfer of a sialic acid from a GD3 oligosaccharide to a different GM3 oligosaccharide (3'-sialyllactoside). The donor substrate specificity study of the CstIIDelta32(I53S) GD3 oligosaccharide synthase activity indicates that the enzyme is flexible in using different CMP-activated sialic acids and their analogs for the synthesis of GD3 oligosaccharides containing natural and nonnatural modifications at the terminal sialic acid.
Department of Chemistry, University of California, One Shields Avenue, Davis, CA 95616, USA.
Jiansong
J
Cheng
Hai
H
Yu
Kam
K
Lau
Shengshu
S
Huang
Harshal A
HA
Chokhawala
Yanhong
Y
Li
Vinod Kumar
VK
Tiwari
Xi
X
Chen
D000595:Amino Acid Sequence; D001483:Base Sequence; D016123:Campylobacter jejuni / Q000201:enzymology* / Q000235:genetics; D003001:Cloning, Molecular; D005732:Gangliosides / Q000378:metabolism; D006023:Glycoproteins / Q000378:metabolism*; D007700:Kinetics; D007790:Lactosylceramides / Q000378:metabolism; D008969:Molecular Sequence Data; D009439:Neuraminidase / Q000378:metabolism*; D012799:Sialyltransferases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism / Q000502:physiology*; D013379:Substrate Specificity
D005732:Gangliosides; D006023:Glycoproteins; D007790:Lactosylceramides; C026226:ganglioside, GD3; C047989:trisialosyllactosylceramide; D012799:Sialyltransferases; C043662:alpha-N-acetylneuraminate alpha-2,8-sialyltransferase; C069715:trans-sialidase; D009439:Neuraminidase