Chemoenzymatic synthesis of stable isotope labeled UDP-N-[2H]-acetyl-glucosamine and [2H]-acetyl-chitooligosaccharides.
Labeled UDP-GlcNAc and chitooligosaccharides should be powerful tools for studies of N-acetylglucosaminyltransferase such as chitin synthases. We describe here a rapid, inexpensive and a common strategie for the chemoenzymatic synthesis of uridine 5'-diphospho-N-[(2)H]-acetyl-glucosamine and the chemical preparation of N-[(2)H]-acetyl chitooligosaccharides (from 2 to 5 mers). Deuterated UDP-GlcNAc analogue was tested as chitin synthase substrate and it exhibited an incorporation level in chitin as the natural substrate. Deuterium labeling of carbohydrates present different advantages: it is a stable isotope and allows glycosyltransferase mechanism studies by a mass spectrometry approach.
Becker
Hubert F
HF
Synthèse, Structure et Fonction de Molécules Bioactives UMR7613, Université Pierre et Marie Curie, Tour 44-45, 3ème étage, 4 place Jussieu, 75252, Paris, France. hbecker@moka.ccr.jussieu.fr
Thellend
Annie
A
Piffeteau
Annie
A
Vidal-Cros
Anne
A
D000107:Acetylation; D002240:Carbohydrate Sequence; D002686:Chitin / Q000378:metabolism*; D002687:Chitin Synthase / Q000032:analysis / Q000378:metabolism; D002855:Chromatography, Thin Layer; D003903:Deuterium / Q000378:metabolism*; D015394:Molecular Structure; D009844:Oligosaccharides / Q000378:metabolism*; D021241:Spectrometry, Mass, Electrospray Ionization; D014537:Uridine Diphosphate N-Acetylglucosamine / Q000138:chemical synthesis* / Q000737:chemistry
D009844:Oligosaccharides; D002686:Chitin; D014537:Uridine Diphosphate N-Acetylglucosamine; D003903:Deuterium; D002687:Chitin Synthase