Synthesis of unnatural sugar nucleotides and their evaluation as donor substrates in glycosyltransferase-catalyzed reactions.
New unnatural sugar nucleotides, UDP-Fuc and CDP-Fuc were synthesized from fucose-beta-1-phosphate and nucleotide monophosphates activated as morpholidates. Furthermore, a nucleotide analogue was prepared by phosphorylation of 1-(beta-D-ribofuranosyl)cyanuric acid, itself obtained as a protected derivative by condensation of the persilylated derivative of cyanuric acid with 1-O-acetyl-2,3,5-tri-O-benzoyl-beta-D-ribofuranose in 74% yield. This phosphate activated according to the same procedure was condensed with fucose-beta-1-phosphate, affording a new sugar nucleotide conjugate (NDP-Fuc) which was evaluated together with UDP-Fuc, CDP-Fuc and ADP-Fuc, as fucose donors in alpha-(1-->4/3)-fucosyltransferase (FucT-III) catalyzed reaction. Fucose transfer could be observed with each of the donors and kinetic parameters were determined using a fluorescent acceptor substrate. Efficiency of the four analogues towards FucT-III was in the following order: UDP-Fuc=ADP-Fuc>NDP-Fuc>CDP-Fuc. According to the same strategy ADP-GlcNAc was prepared from AMP-morpholidate and N-acetylglucosamine-alpha-1-phosphate; tested as a glucosaminyl donor towards Neisseria meningitidis N-acetylglucosaminyl transferase (LgtA), ADP-GlcNAc was recognized with 0.1% efficiency as compared with UDP-GlcNAc, the natural donor substrate.
Khaled
Amira
A
Laboratoire de Chimie Organique Multifonctionnelle, UMR 8614, GDR 2590, Institut de Chimie Moléculaire et des Matériaux d'Orsay (ICMMO), Université Paris-Sud, Bât 420, F-91405 Orsay, France.
Ivannikova
Tatiana
T
Augé
Claudine
C
D000247:Adenosine Diphosphate Sugars / Q000138:chemical synthesis / Q000378:metabolism; D001426:Bacterial Proteins / Q000378:metabolism; D002384:Catalysis; D005647:Fucosyltransferases / Q000378:metabolism; D016695:Glycosyltransferases / Q000378:metabolism*; D007700:Kinetics; D017351:N-Acetylglucosaminyltransferases / Q000378:metabolism; D009702:Nucleoside Diphosphate Sugars / Q000138:chemical synthesis* / Q000378:metabolism; D013329:Structure-Activity Relationship; D013379:Substrate Specificity; D014539:Uridine Diphosphate Sugars / Q000138:chemical synthesis / Q000378:metabolism
D000247:Adenosine Diphosphate Sugars; D001426:Bacterial Proteins; D009702:Nucleoside Diphosphate Sugars; D014539:Uridine Diphosphate Sugars; D016695:Glycosyltransferases; D005647:Fucosyltransferases; C401409:LgtA protein, bacteria; D017351:N-Acetylglucosaminyltransferases; C019818:3-galactosyl-N-acetylglucosaminide 4-alpha-L-fucosyltransferase