Synthesis of sugar arrays in microtiter plate.
1,3-Dipolar cycloadditions between azides and alkynes were exploited to attach oligosaccharides to a C(14) hydrocarbon chain that noncovalently binds to the microtiter well surface. Synthesis of sugar arrays was performed on a micromolar scale in situ in the microtiter plate. As a model study, the beta-galactosyllipid 5 was displayed on a 4-micromol scale. Formation of product was confirmed via ESI-MS, and the yield was determined via chemical and biological assays. Several complex carbohydrates (6-16) were also displayed in microtiter plates and successfully screened with various lectins. Moreover, sialyl Lewis x (17) was synthesized via the enzymatic fucosylation of a precursor displayed in the plate. Studies on inhibition of this biotransformation have been carried out, and the IC(50) value found for the known inhibitor 20 was consistent with previous studies in solution.
Fazio
Fabio
F
Department of Chemistry and Skaggs Institute for Chemical Biology, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, California 92037, USA.
Bryan
Marian C
MC
Blixt
Ola
O
Paulson
James C
JC
Wong
Chi-Huey
CH
D000469:Alkaline Phosphatase / Q000737:chemistry; D000480:Alkynes / Q000737:chemistry; D000911:Antibodies, Monoclonal / Q000737:chemistry; D001386:Azides / Q000737:chemistry; D002240:Carbohydrate Sequence; D008969:Molecular Sequence Data; D019906:Nuclear Magnetic Resonance, Biomolecular; D009844:Oligosaccharides / Q000138:chemical synthesis* / Q000737:chemistry; D037121:Plant Lectins / Q000737:chemistry; D054788:Ribosome Inactivating Proteins; D012276:Ricin / Q000737:chemistry; D012710:Serum Albumin, Bovine / Q000737:chemistry; D000080506:Sialyl Lewis X Antigen; D021241:Spectrometry, Mass, Electrospray Ionization
D000480:Alkynes; D000911:Antibodies, Monoclonal; D001386:Azides; D009844:Oligosaccharides; D037121:Plant Lectins; C050600:Sambucus nigra lectins; D000080506:Sialyl Lewis X Antigen; D012710:Serum Albumin, Bovine; D012276:Ricin; D000469:Alkaline Phosphatase; D054788:Ribosome Inactivating Proteins