title: Expression, characterization and homology modeling of a novel eukaryotic GH84 β-N-acetylglucosaminidase from Penicillium chrysogenum.
abstract: 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.
authors.affiliation: Laboratory of Biotransformation, Institute of Microbiology, Academy of Sciences of the Czech Republic, Vídeňská 1083, 14220 Praha 4, Czech Republic. Electronic address: slamova@biomed.cas.cz.
authors.forename: Kristýna
authors.initials: K
authors.lastname: Slámová
authors.affiliation: Department of Structure and Function of Proteins, Institute of Nanobiology and Structural Biology of GCRC, Zámek 136, 37333 Nové Hrady, Czech Republic.
authors.forename: Natallia
authors.initials: N
authors.lastname: Kulik
authors.affiliation: Laboratory of Biotransformation, Institute of Microbiology, Academy of Sciences of the Czech Republic, Vídeňská 1083, 14220 Praha 4, Czech Republic.
authors.forename: Martin
authors.initials: M
authors.lastname: Fiala
authors.affiliation: Laboratory of Biotransformation, Institute of Microbiology, Academy of Sciences of the Czech Republic, Vídeňská 1083, 14220 Praha 4, Czech Republic; Department of Biochemistry and Microbiology, Institute of Chemical Technology Prague, Technická 5, 16628 Praha 6, Czech Republic.
authors.forename: Jana
authors.initials: J
authors.lastname: Krejzová-Hofmeisterová
authors.affiliation: Department of Structure and Function of Proteins, Institute of Nanobiology and Structural Biology of GCRC, Zámek 136, 37333 Nové Hrady, Czech Republic.
authors.forename: Rüdiger
authors.initials: R
authors.lastname: Ettrich
authors.affiliation: Laboratory of Biotransformation, Institute of Microbiology, Academy of Sciences of the Czech Republic, Vídeňská 1083, 14220 Praha 4, Czech Republic.
authors.forename: Vladimír
authors.initials: V
authors.lastname: Křen
mesh_terms: D000118:Acetylglucosaminidase / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D000595:Amino Acid Sequence; D062105:Molecular Docking Simulation; D008969:Molecular Sequence Data; D010408:Penicillium chrysogenum / Q000201:enzymology / Q000235:genetics*; D010843:Pichia / Q000235:genetics / Q000378:metabolism; D011485:Protein Binding; D011994:Recombinant Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D016415:Sequence Alignment
chemical_list: D011994:Recombinant Proteins; D000118:Acetylglucosaminidase
keywords: Homology modeling; O-GlcNAcase; Pichia pastoris; Yeast expression system; β-N-acetylglucosaminidase