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Taxonomic group: fungi / Ascomycota (Phylum: Ascomycota)

The structure was elucidated in this paper
NCBI PubMed ID: 29239122
Publication DOI: 10.1111/febs.14360
Journal NLM ID: 101229646
Publisher: Blackwell Publishing
Correspondence: Vaněk O <ondrej.vanek

natur.cuni.cz>; Řezáčová P <rezacova

uochb.cas.cz>
Institutions: Institute of Organic Chemistry and Biochemistry, The Czech Academy of Sciences, Prague, Czech Republic, Institute of Molecular Genetics, The Czech Academy of Sciences, Prague, Czech Republic, Department of Biochemistry, Faculty of Science, Charles University, Prague, Czech Republic, Institute of Microbiology, The Czech Academy of Sciences, Prague, Czech Republic, Institute of Physics, Faculty of Mathematics and Physics, Charles University, Prague, Czech Republic, UT Southwestern Medical Center, Dallas, USA

β-N-acetylhexosaminidase from the fungus Aspergillus oryzae is a secreted extracellular enzyme that cleaves chitobiose into constituent monosaccharides. It belongs to the GH 20 glycoside hydrolase family and consists of two N-glycosylated catalytic cores noncovalently associated with two 10-kDa O-glycosylated propeptides. We used X-ray diffraction and mass spectrometry to determine the structure of A. oryzae β-N-acetylhexosaminidase isolated from its natural source. The three-dimensional structure determined and refined to a resolution of 2.3 Å revealed that this enzyme is active as a uniquely tight dimeric assembly further stabilized by N- and O-glycosylation. The propeptide from one subunit forms extensive noncovalent interactions with the catalytic core of the second subunit in the dimer, and this chain swap suggests the distinctive structural mechanism of the enzyme's activation. Unique structural features of β-N-acetylhexosaminidase from A. oryzae define a very stable and robust framework suitable for biotechnological applications. The crystal structure reported here provides structural insights into the enzyme architecture as well as the detailed configuration of the active site. These insights can be applied to rational enzyme engineering.

mass spectrometry, glycosylation, X-ray crystallography, active pocket, carbohydrate biotechnology, propeptide

Structure type: monomer
Location inside paper: p. 584, column 2, paragraph 2
Aglycon: (->3) L-Ser/L-Thr (AoHEX)
Compound class: O-glycan
Contained glycoepitopes: IEDB_137485,IEDB_144983,IEDB_152206,IEDB_983930,SB_44,SB_72

Methods: gel filtration, SDS-PAGE, MS/MS, LC-MS, reversed-phase chromatography, precipitation, x-ray, FTIR-MS
Comments, role: N-glycan moiety attached to T78, S83, S84; stabilizes the structure of protein

Related record ID(s): 49073, 49074, 49152
NCBI Taxonomy refs (TaxIDs): 5062
Reference(s) to other database(s): GTC:G74840NI
Show glycosyltransferases

There is only one chemically distinct structure:

OC[C@H]1O[C@@H](O)[C@@H](O)[C@@H](O)[C@@H]1O

SVG MW SMILES 3D mol Ionize

natur.cuni.cz>; Řezáčová P <rezacova

mass spectrometry, glycosylation, X-ray crystallography, active pocket, carbohydrate biotechnology, propeptide

Structure type: oligomer
Location inside paper: p. 4, column 2, table 2 line 6, fig. 3(B)
Aglycon: (->4) Asn-X-Ser/Thr (AoHEX)
Compound class: N-glycan
Contained glycoepitopes: IEDB_135813,IEDB_137340,IEDB_137485,IEDB_141807,IEDB_144983,IEDB_151531,IEDB_152206,IEDB_153212,IEDB_983930,SB_44,SB_72,SB_74,SB_85

Methods: gel filtration, SDS-PAGE, MS/MS, LC-MS, reversed-phase chromatography, precipitation, x-ray, FTIR-MS
Synthetic data: enzymatic in vivo
Comments, role: N-glycan moiety attached to Asn353; stabilizes the structure of protein

Related record ID(s): 49055, 49074, 49152
NCBI Taxonomy refs (TaxIDs): 5062
Reference(s) to other database(s): GTC:G15407YE
Show glycosyltransferases

There is only one chemically distinct structure:

CC(=O)N[C@H]1[C@H](O[C@@H]2[C@@H](CO)O[C@@H](O)[C@H](NC(C)=O)[C@H]2O)O[C@H](CO)[C@@H](O[C@@H]2O[C@H](CO)[C@@H](O)[C@H](O)[C@@H]2O)[C@@H]1O

SVG MW SMILES 3D mol Ionize

natur.cuni.cz>; Řezáčová P <rezacova

mass spectrometry, glycosylation, X-ray crystallography, active pocket, carbohydrate biotechnology, propeptide

Structure type: oligomer
Location inside paper: p. 4, column 2, table 2 line 8
Aglycon: (->4) Asn-X-Ser/Thr (AoHEX)
Compound class: N-glycan
Contained glycoepitopes: IEDB_135813,IEDB_137340,IEDB_141807,IEDB_151531,IEDB_153212,SB_74,SB_85

Methods: gel filtration, SDS-PAGE, MS/MS, LC-MS, reversed-phase chromatography, precipitation, x-ray, FTIR-MS
Synthetic data: enzymatic in vivo
Comments, role: N-glycan moiety attached to Asn428; stabilizes the structure of protein

Related record ID(s): 49055, 49073, 49152
NCBI Taxonomy refs (TaxIDs): 5062
Reference(s) to other database(s): GTC:G42666HT
Show glycosyltransferases

There is only one chemically distinct structure:

CC(=O)N[C@H]1[C@H](O[C@@H]2[C@@H](CO)O[C@@H](O)[C@H](NC(C)=O)[C@H]2O)O[C@H](CO)[C@@H](O)[C@@H]1O

SVG MW SMILES 3D mol Ionize

natur.cuni.cz>; Řezáčová P <rezacova

mass spectrometry, glycosylation, X-ray crystallography, active pocket, carbohydrate biotechnology, propeptide

Structure type: monomer
Location inside paper: p. 4, column 2, table 2 line 9
Aglycon: (->4) Asn-X-Ser/Thr (AoHEX)
Compound class: N-glycan
Contained glycoepitopes: IEDB_135813,IEDB_137340,IEDB_141807,IEDB_151531

Methods: gel filtration, SDS-PAGE, MS/MS, LC-MS, reversed-phase chromatography, precipitation, x-ray, FTIR-MS
Synthetic data: enzymatic in vivo
Comments, role: N-glycan moiety attached to Asn500; stabilizes the structure of protein

Related record ID(s): 49055, 49073, 49074
NCBI Taxonomy refs (TaxIDs): 5062
Reference(s) to other database(s): GTC:G49108TO
Show glycosyltransferases

There is only one chemically distinct structure:

CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O

SVG MW SMILES 3D mol Ionize