Expression, characterization and homology modeling of a novel eukaryotic GH84 β-N-acetylglucosaminidase from Penicillium chrysogenum
| Autor: | Martin Fiala, Rüdiger Ettrich, Vladimír Křen, Jana Krejzová-Hofmeisterová, Natallia Kulik, Kristýna Slámová |
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| Rok vydání: | 2013 |
| Předmět: |
Cell
Molecular Sequence Data Penicillium chrysogenum 010402 general chemistry 01 natural sciences Pichia Pichia pastoris 03 medical and health sciences N acetylglucosaminidase Acetylglucosaminidase medicine Glycoside hydrolase Homology modeling Amino Acid Sequence 030304 developmental biology 0303 health sciences biology biology.organism_classification Recombinant Proteins 0104 chemical sciences Molecular Docking Simulation Cytosol medicine.anatomical_structure Biochemistry Docking (molecular) Sequence Alignment Biotechnology Protein Binding |
| Zdroj: | Protein Expression and Purification |
| ISSN: | 1096-0279 |
| Popis: | β-N-acetylglucosaminidases from the family 84 of glycoside hydrolases form a small group of glycosidases in eukaryotes responsible for the modification of nuclear and cytosolic proteins with O-GlcNAc, thus they are involved in a number of important cell processes. Here, the first fungal β-N-acetylglucosaminidase from Penicillium chrysogenum was expressed in Pichia pastoris and secreted into the media, purified and characterized. Moreover, homology modeling and substrate and inhibitor docking were performed to obtain structural information on this new member of the GH84 family. Surprisingly, we found that this fungal β-N-acetylglucosaminidase with its sequence and structure perfectly fitting to the GH84 family displays biochemical properties rather resembling the β-N-acetylhexosaminidases from the family 20 of glycoside hydrolases. This work helped to increase the knowledge on the scarcely studied glycosidase family and revealed a new type of eukaryotic β-N-acetylglucosaminidase. |
| Databáze: | OpenAIRE |
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