Biomaterial Database

Structural basis and catalytic mechanism for the dual functional endo-beta-N-acetylglucosaminidase A. 2009

Jie Yin, and Lei Li, and Neil Shaw, and Yang Li, and Jing Katherine Song, and Wenpeng Zhang, and Chengfeng Xia, and Rongguang Zhang, and Andrzej Joachimiak, and Hou-Cheng Zhang, and Lai-Xi Wang, and Zhi-Jie Liu, and Peng Wang

National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing, China.

Endo-beta-N-acetylglucosaminidases (ENGases) are dual specificity enzymes with an ability to catalyze hydrolysis and transglycosylation reactions. Recently, these enzymes have become the focus of intense research because of their potential for synthesis of glycopeptides. We have determined the 3D structures of an ENGase from Arthrobacter protophormiae (Endo-A) in 3 forms, one in native form, one in complex with Man(3)GlcNAc-thiazoline and another in complex with GlcNAc-Asn. The carbohydrate moiety sits above the TIM-barrel in a cleft region surrounded by aromatic residues. The conserved essential catalytic residues - E173, N171 and Y205 are within hydrogen bonding distance of the substrate. W216 and W244 regulate access to the active site during transglycosylation by serving as "gate-keepers". Interestingly, Y299F mutation resulted in a 3 fold increase in the transglycosylation activity. The structure provides insights into the catalytic mechanism of GH85 family of glycoside hydrolases at molecular level and could assist rational engineering of ENGases.

UI	MeSH Term	Description	Entries
D008958	Models, Molecular	Models used experimentally or theoretically to study molecular shape, electronic properties, or interactions; includes analogous molecules, computer-generated graphics, and mechanical structures.	Molecular Models,Model, Molecular,Molecular Model
D008969	Molecular Sequence Data	Descriptions of specific amino acid, carbohydrate, or nucleotide sequences which have appeared in the published literature and/or are deposited in and maintained by databanks such as GENBANK, European Molecular Biology Laboratory (EMBL), National Biomedical Research Foundation (NBRF), or other sequence repositories.	Sequence Data, Molecular,Molecular Sequencing Data,Data, Molecular Sequence,Data, Molecular Sequencing,Sequencing Data, Molecular
D011487	Protein Conformation	The characteristic 3-dimensional shape of a protein, including the secondary, supersecondary (motifs), tertiary (domains) and quaternary structure of the peptide chain. PROTEIN STRUCTURE, QUATERNARY describes the conformation assumed by multimeric proteins (aggregates of more than one polypeptide chain).	Conformation, Protein,Conformations, Protein,Protein Conformations
D002240	Carbohydrate Sequence	The sequence of carbohydrates within POLYSACCHARIDES; GLYCOPROTEINS; and GLYCOLIPIDS.	Carbohydrate Sequences,Sequence, Carbohydrate,Sequences, Carbohydrate
D013379	Substrate Specificity	A characteristic feature of enzyme activity in relation to the kind of substrate on which the enzyme or catalytic molecule reacts.	Specificities, Substrate,Specificity, Substrate,Substrate Specificities
D016297	Mutagenesis, Site-Directed	Genetically engineered MUTAGENESIS at a specific site in the DNA molecule that introduces a base substitution, or an insertion or deletion.	Mutagenesis, Oligonucleotide-Directed,Mutagenesis, Site-Specific,Oligonucleotide-Directed Mutagenesis,Site-Directed Mutagenesis,Site-Specific Mutagenesis,Mutageneses, Oligonucleotide-Directed,Mutageneses, Site-Directed,Mutageneses, Site-Specific,Mutagenesis, Oligonucleotide Directed,Mutagenesis, Site Directed,Mutagenesis, Site Specific,Oligonucleotide Directed Mutagenesis,Oligonucleotide-Directed Mutageneses,Site Directed Mutagenesis,Site Specific Mutagenesis,Site-Directed Mutageneses,Site-Specific Mutageneses
D017038	Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase	A group of related enzymes responsible for the endohydrolysis of the di-N-acetylchitobiosyl unit in high-mannose-content glycopeptides and GLYCOPROTEINS.	Di-N-Acetylchitobiosyl beta-N-Acetylglucosaminidase,Endo-beta-Acetylglucosaminidase,Endoglycosidase F,Endo D Endoglucosaminidase,Endo F Endoglucosaminidase,Endo H Endoglucosaminidase,Endo-N-Acetyl-beta-d-glucosaminidase,Endo-beta-N-Acetylglucosaminidase D,Endo-beta-N-Acetylglucosaminidase F,Endo-beta-N-Acetylglucosaminidase H,Endoglucosaminidase D,Endoglucosaminidase F,Endoglucosaminidase H,Endoglucosidase H,Endoglycosidase D,Endohexosaminidase F,Endohexosaminidase H,Peptide N-Glycosidase F,Di N Acetylchitobiosyl beta N Acetylglucosaminidase,Endo N Acetyl beta d glucosaminidase,Endo beta Acetylglucosaminidase,Endo beta N Acetylglucosaminidase D,Endo beta N Acetylglucosaminidase F,Endo beta N Acetylglucosaminidase H,Endo-beta-N-Acetylglucosaminidase, Mannosyl-Glycoprotein,Mannosyl Glycoprotein Endo beta N Acetylglucosaminidase,Peptide N Glycosidase F,beta-N-Acetylglucosaminidase, Di-N-Acetylchitobiosyl
D055162	Biocatalysis	The facilitation of biochemical reactions with the aid of naturally occurring catalysts such as ENZYMES.