Optimized deglycosylation of glycoproteins by peptide-N4-(N-acetyl-β-glucosaminyl)-asparagine amidase fromFlavobacterium meningosepticum
| العنوان: | Optimized deglycosylation of glycoproteins by peptide-N4-(N-acetyl-β-glucosaminyl)-asparagine amidase fromFlavobacterium meningosepticum |
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| المؤلفون: | Martin Zimmermann, Rolf Nuck, D Sauvageot, Josi D, Werner Reutter |
| المصدر: | Glycoconjugate Journal. 7:279-286 |
| بيانات النشر: | Springer Science and Business Media LLC, 1990. |
| سنة النشر: | 1990 |
| مصطلحات موضوعية: | chemistry.chemical_classification, PNGase F, Glycosylation, Chromatography, biology, Chemistry, Peptide, Cell Biology, biology.organism_classification, Flavobacterium, Biochemistry, Amidohydrolases, Amidase, chemistry.chemical_compound, Enzyme, Humans, Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase, Asparagine, Glycoprotein, Molecular Biology, Chromatography, High Pressure Liquid, Glycoproteins |
| الوصف: | Peptide-N4-(N-acetyl-beta-glucosaminyl)asparagine amidase F(PNGase F) from Flavobacterium meningosepticum is a highly useful enzyme for the structural analysis of N (asparagine)-linked carbohydrate chains derived from glycoproteins. The enzyme was enriched using a published procedure [Tarentino AL, Gomez CM, Plummer TH, Jr (1984) Biochemistry 1985:4665-71; Tarentino AL, Plummer TH, Jr (1987) Methods Enzymol 138:770-78] and further purified by hydrophobic interaction HPLC on a weak hydrophobic TSK-Ether column from which it was eluted by a decreasing gradient of 1.7 M ammonium sulphate in 100 mM sodium phosphate, pH 7.0, containing 5 mM EDTA. To determine the optimal conditions for a complete deglycosylation of glycoproteins by PNGase F, experiments were performed with human alpha 1-acid glycoprotein, because the five complex type carbohydrate chains are quite resistant to enzymic hydrolysis. The influence of different detergents on the enzyme reaction was studied. Complete deglycosylation of human alpha 1-acid glycoprotein was achieved by the use of 60 mU/ml PNGase F in 0.25 M sodium phosphate buffer, pH 8.6, containing 0.2% (w/v) SDS, 20 mM mercaptoethanol and 0.5% Mega-10. |
| تدمد: | 1573-4986 0282-0080 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::f10a66ece25993fdf12004a1bfa034e1 https://doi.org/10.1007/bf01073372 |
| حقوق: | CLOSED |
| رقم الأكسشن: | edsair.doi.dedup.....f10a66ece25993fdf12004a1bfa034e1 |
| قاعدة البيانات: | OpenAIRE |
Find this article in full text from Springer Verlag. | تدمد: | 15734986 02820080 |
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