Characterization of the peptide-N4-(N-acetylglucosaminyl) asparagine amidase (PNGase Se) fromSilene alba cells |
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Authors: | Sabine Lhernould Yannis Karamanos Patrice Lerouge and Henri Morvan |
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Institution: | (1) Laboratoire de Biologie Cellulaire Végétale et Valorisation des Espéces Ligneuses, Université de Limoges, 123, Avenue Albert Thomas, 87060 Limoges Cédex, France;(2) Institut de Biotechnologie, Université de Limoges, 123, Avenue Albert Thomas, 87060 Limoges Cédex, France;(3) Laboratoire des transports Intracellulaires, URA 203, UFR des Sciences, Université de Rouen, 76821 Mont St Aignan Cédex, France |
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Abstract: | The peptide-N
4-(N-acetylglucosaminyl) asparagine amidase (PNGase Se) earlier described Lhernould S., Karamanos Y., Bourgerie S., Strecker G., Julien R., Morvan H. (1992)Glycoconjugate J
9:191–97] was partially purified from culturedSilene alba cells using affinity chromatography. The enzyme is active between pH 3.0 and 6.5, and is stable in the presence of moderate concentrations of several other protein unfolding chemicals, but is readily inactivated by SDS. Although the enzyme cleaves the carbohydrate from a variety of animal and plant glycopeptides, it does not hydrolyse the carbohydrate from most of the corresponding unfolded glycoproteins in otherwise comparable conditions. The substrate specificity of this plant PNGase supports the hypothesis that this enzyme could be at the origin of the production of unconjugated N-glycans in a suspension medium of culturedSilene alba cells.Abbreviations GlcNAc
N-acetylglucosamine
- PNGase
peptide-N
4-(N-acetylglucosaminyl) asparagine amidase
- BSA
bovine serum albumin
- SDS-PAGE
sodium dodecyl sulfate-polyacrylamide gel electrophoresis
- TLC
thin layer chromatography
- HPAEC-PAD
High pH anion exchange chromatography-pulsed amperometric detection |
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Keywords: | de-N-glycosylation PNGase white campion |
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