Versatile Glycoblotting Nanoparticles for High-Throughput Protein Glycomics
| Autor: | Hirosato Kondo, Hiroaki Nakagawa, Kenji Monde, Kisaburo Deguchi, Shin-Ichiro Nishimura, Kenichi Niikura, Masaki Kurogochi, Yasuro Shinohara, Ryosuke Kamitani, Rie Uematsu |
|---|---|
| Rok vydání: | 2005 |
| Předmět: |
PNGase F
Chromatography Time Factors Aqueous solution Glycosylation Photochemistry High-throughput screening Organic Chemistry Glycopeptides Oligosaccharides Nanoparticle General Chemistry Mass spectrometry Catalysis Nanostructures Glycomics Mice chemistry.chemical_compound chemistry Spectrometry Mass Matrix-Assisted Laser Desorption-Ionization Liposomes Animals Centrifugation Skin |
| Zdroj: | Chemistry - A European Journal. 11:3825-3834 |
| ISSN: | 1521-3765 0947-6539 |
| Popis: | We have developed an effective and practical trap-and-release method based on chemoselective ligation of carbohydrates with reactive aminooxyl groups attached to the surface of nanoparticles (referred to as glycoblotting nanoparticles). These glycoblotting nanoparticles were synthesized by UV irradiation of diacetylene-functionalized lipids that contain the aminooxyl group. The glycoblotting nanoparticles captured carbohydrates in aqueous solution under mild conditions and were collected by simple centrifugation. The trapped carbohydrates were effectively released from the nanoparticles under acidic conditions to give pure oligosaccharides. This glycoblotting process reduced the time required for the purification process of carbohydrates to less than 6 h, compared to the several days needed for conventional chromatographic techniques. The oligosaccharides (N-glycan) were released from ovalbumin (glycoprotein) by PNGase F after tryptic digestion. MALDI-TOF mass spectra before purification did not show any significant signals corresponding to N-glycans because these signals were hidden by the large signals of the abundant peptides. However, after purification with the glycoblotting nanoparticles, only signals corresponding to oligosaccharides appeared. We also demonstrated a clear analysis of the oligosaccharides contained in the mice dermis by means of glycoblotting. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Plný text