Chemoenzymatic synthesis of GDP-L-fucose and the Lewis X glycan derivatives.
Lewis X (Le(x))-containing glycans play important roles in numerous cellular processes. However, the absence of robust, facile, and cost-effective methods for the synthesis of Le(x) and its structurally related analogs has severely hampered the elucidation of the specific functions of these glycan epitopes. Here we demonstrate that chemically defined guanidine 5'-diphosphate-beta-l-fucose (GDP-fucose), the universal fucosyl donor, the Le(x) trisaccharide, and their C-5 substituted derivatives can be synthesized on preparative scales, using a chemoenzymatic approach. This method exploits l-fucokinase/GDP-fucose pyrophosphorylase (FKP), a bifunctional enzyme isolated from Bacteroides fragilis 9343, which converts l-fucose into GDP-fucose via a fucose-1-phosphate (Fuc-1-P) intermediate. Combining the activities of FKP and a Helicobacter pylori alpha1,3 fucosyltransferase, we prepared a library of Le(x) trisaccharide glycans bearing a wide variety of functional groups at the fucose C-5 position. These neoglycoconjugates will be invaluable tools for studying Le(x)-mediated biological processes.
Wang
Wei
W
Department of Biochemistry and Macromolecular Therapeutics Development Facility, Albert Einstein College of Medicine, Yeshiva University, 1300 Morris Park Avenue, Bronx, NY 10461, USA.
Hu
Tianshun
T
Frantom
Patrick A
PA
Zheng
Tianqing
T
Gerwe
Brian
B
Del Amo
David Soriano
DS
Garret
Sarah
S
Seidel
Ronald D
RD
Wu
Peng
P
D000255:Adenosine Triphosphate / Q000378:metabolism; D001426:Bacterial Proteins / Q000378:metabolism*; D001441:Bacteroides fragilis / Q000201:enzymology; D002384:Catalysis; D018360:Crystallography, X-Ray; D004591:Electrophoresis, Polyacrylamide Gel; D006154:Guanosine Diphosphate Fucose / Q000096:biosynthesis* / Q000737:chemistry; D006160:Guanosine Triphosphate / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D016256:Lewis X Antigen / Q000737:chemistry; D009682:Magnetic Resonance Spectroscopy; D015394:Molecular Structure; D009713:Nucleotidyltransferases / Q000378:metabolism*; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000378:metabolism*; D011134:Polysaccharides / Q000096:biosynthesis* / Q000737:chemistry; D013379:Substrate Specificity
D001426:Bacterial Proteins; D016256:Lewis X Antigen; D011134:Polysaccharides; D006154:Guanosine Diphosphate Fucose; D006160:Guanosine Triphosphate; D000255:Adenosine Triphosphate; D017853:Phosphotransferases (Alcohol Group Acceptor); C020153:fucokinase; C115708:GDP-L-fucose pyrophosphorylase; D009713:Nucleotidyltransferases