Transfer of modified sialic acids by Trypanosoma cruzi trans-sialidase for attachment of functional groups to oligosaccharide.
Donor specificity of Trypanosoma cruzi trans-sialidase (TcTs) has been investigated with modified 2-[4-methylumbelliferone]-alpha- ketoside of N-acetyl-D-neuraminic acid (4MU-NANA) as donor and lactose as acceptor. 4MU-NANA was treated with periodate under mild conditions to generate an aldehyde on the exocyclic side chain. The oxidized 4MU-NANA was derivatized with various primary amines by reductive amination to yield potential donors. High-performance anion-exchange chromatography equipped with pulsed amperometric detector was used to assay the transglycosylation activity of TcTs. Several modified 4MU-NANA derivatives served as substrates by TcTs and they may be utilized to make valuable intermediates, including those for fluorescence energy transfer measurement or photoaffinity labeling experiment.
Lee
K B
KB
Department of Biology, Johns Hopkins University, Baltimore, Maryland 21218.
Lee
Y C
YC
D000586:Amination; D000818:Animals; D001426:Bacterial Proteins; D002240:Carbohydrate Sequence; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange / Q000379:methods; D005340:Fibrinogen / Q000378:metabolism; D005456:Fluorescent Dyes / Q000378:metabolism; D006020:Glycopeptides / Q000378:metabolism; D006923:Hymecromone / Q000031:analogs & derivatives / Q000378:metabolism; D008969:Molecular Sequence Data; D009439:Neuraminidase / Q000378:metabolism*; D009844:Oligosaccharides / Q000378:metabolism*; D010084:Oxidation-Reduction; D010504:Periodic Acid; D012794:Sialic Acids / Q000378:metabolism*; D013379:Substrate Specificity; D014349:Trypanosoma cruzi / Q000201:enzymology*
D001426:Bacterial Proteins; D005456:Fluorescent Dyes; D006020:Glycopeptides; D009844:Oligosaccharides; D012794:Sialic Acids; D010504:Periodic Acid; D006923:Hymecromone; C021224:2'-(4-methylumbelliferyl)-alpha-D-N-acetylneuraminic acid; D005340:Fibrinogen; D009439:Neuraminidase