Novel fluorescent asparaginyl-N-acetyl-d-glucosamines (Asn-GlcNAc) for the resolution of oligosaccharides in glycopeptides, based on enzyme transglycosylation reaction
| Autor: | Hiroki Kawanishi, Jun Zhe Min, Masaru Kato, Takeshi Fukushima, Toshimasa Toyo'oka |
|---|---|
| Rok vydání: | 2005 |
| Předmět: |
chemistry.chemical_classification
biology Stereochemistry Carboxylic acid Oligosaccharide biology.organism_classification Biochemistry Acceptor Fluorescence Fluorescence spectroscopy Analytical Chemistry carbohydrates (lipids) Residue (chemistry) Mucor hiemalis chemistry Reagent Environmental Chemistry Organic chemistry Spectroscopy |
| Zdroj: | Analytica Chimica Acta. 550:173-181 |
| ISSN: | 0003-2670 |
| Popis: | Several fluorescent asparaginyl- N -acetyl- d -glucosamines (Asn-GlcNAcs), i.e., DBD-Asn-GlcNAc, NBD-Asn-GlcNAc, NDA-Asn-GlcNAc, PS-Asn-GlcNAc, FITC-Asn-GlcNAc, DMEQ-Asn-GlcNAc and DBD-PZ-Boc-Asn-GlcNAc, were synthesized as the acceptors for the resolution of oligosaccharides in glycopeptides. The resolution of oligosaccharides is based on the transglycosylation reaction with end-β- N -acetylglucosaminidase (Endo-M), isolated from Mucor hiemalis . The synthesis of each fluorescent acceptor was carried out in a one-pot reaction of Asn-GlcNAc and the corresponding fluorescent tagging reagent. The transglycosylation reaction using Endo-M proceeds at RT in neutral phosphate buffer (pH 6.0) and reached maxima at around 30 min. When Fmoc-Asn-GlcNAc (acceptor) was enzymatically reacted with Disialo-Asn (donor) in the presence of Endo-M, the ratio of Disialo-Asn-Fmoc (transglycosylation product) was almost comparable with the decreasing ratio of Fmoc-Asn-GlcNAc. Therefore, the transglycosylation activity of Endo-M from Disialo-Asn (donor) and fluorescent-Asn-GlcNAc (acceptor) was calculated from the decreasing ratio of fluorescent-Asn-GlcNAc. The order was NDA-Asn-GlcNAc > DBD-Asn-GlcNAc ≅ Fmoc-Asn-GlcNAc > NBD-Asn-GlcNAc ≅ DMEQ-Asn-GlcNAc > DNS-Asn-GlcNAc > PS-Asn-GlcNAc > FITC-Asn-GlcNAc. On the other hand, the activity with a fluorescent acceptor (DBD-PZ-Boc-Asn-GlcNAc), labeled to a carboxylic acid group in the Asn residue, was the strongest among the synthesized acceptors. |
| Databáze: | OpenAIRE |
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