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Identification of the catalytic acid base residue of arthrobacter endo-beta-N-acetylglucosaminidase by chemical rescue of an inactive mutant. 2007

Kiyotaka Fujita, and Reiko Sato, and Kazunori Toma, and Kanefumi Kitahara, and Toshihiko Suganuma, and Kenji Yamamoto, and Kaoru Takegawa
Faculty of Agriculture, Kagoshima University, Korimoto, Kagoshima, Japan. kfujita@ms.kagoshima-u.ac.jp

Arthrobacter endo-beta-N-acetylglucosaminidase (Endo-A), a member of glycoside hydrolase (GH) family 85, catalyses the hydrolysis and transglycosylation of asparagine-linked oligosaccharides of glycoproteins with retention of anomeric configuration. Glu-173 of Endo-A is a catalytically essential amino acid residue, and the corresponding residue is conserved in all GH family 85 members. The catalytic activity of Endo-A E173A mutant was rescued by the addition of sodium azide or sodium formate. Furthermore, the produced beta-glycosyl azide (Man(5)GlcNAc-beta-N(3)) retained the anomeric configuration, indicating that Glu-173 is the catalytic acid-base residue of Endo-A. This is the first identification of the catalytic residue for GH family 85 endo-beta-N-acetylglucosaminidases.

UI MeSH Term Description Entries
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
D009154 Mutation Any detectable and heritable change in the genetic material that causes a change in the GENOTYPE and which is transmitted to daughter cells and to succeeding generations. Mutations
D002240 Carbohydrate Sequence The sequence of carbohydrates within POLYSACCHARIDES; GLYCOPROTEINS; and GLYCOLIPIDS. Carbohydrate Sequences,Sequence, Carbohydrate,Sequences, Carbohydrate
D002384 Catalysis The facilitation of a chemical reaction by material (catalyst) that is not consumed by the reaction. Catalyses
D002851 Chromatography, High Pressure Liquid Liquid chromatographic techniques which feature high inlet pressures, high sensitivity, and high speed. Chromatography, High Performance Liquid,Chromatography, High Speed Liquid,Chromatography, Liquid, High Pressure,HPLC,High Performance Liquid Chromatography,High-Performance Liquid Chromatography,UPLC,Ultra Performance Liquid Chromatography,Chromatography, High-Performance Liquid,High-Performance Liquid Chromatographies,Liquid Chromatography, High-Performance
D004591 Electrophoresis, Polyacrylamide Gel Electrophoresis in which a polyacrylamide gel is used as the diffusion medium. Polyacrylamide Gel Electrophoresis,SDS-PAGE,Sodium Dodecyl Sulfate-PAGE,Gel Electrophoresis, Polyacrylamide,SDS PAGE,Sodium Dodecyl Sulfate PAGE,Sodium Dodecyl Sulfate-PAGEs
D000595 Amino Acid Sequence The order of amino acids as they occur in a polypeptide chain. This is referred to as the primary structure of proteins. It is of fundamental importance in determining PROTEIN CONFORMATION. Protein Structure, Primary,Amino Acid Sequences,Sequence, Amino Acid,Sequences, Amino Acid,Primary Protein Structure,Primary Protein Structures,Protein Structures, Primary,Structure, Primary Protein,Structures, Primary Protein
D001173 Arthrobacter A genus of asporogenous bacteria isolated from soil that displays a distinctive rod-coccus growth cycle.
D001483 Base Sequence The sequence of PURINES and PYRIMIDINES in nucleic acids and polynucleotides. It is also called nucleotide sequence. DNA Sequence,Nucleotide Sequence,RNA Sequence,DNA Sequences,Base Sequences,Nucleotide Sequences,RNA Sequences,Sequence, Base,Sequence, DNA,Sequence, Nucleotide,Sequence, RNA,Sequences, Base,Sequences, DNA,Sequences, Nucleotide,Sequences, RNA
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

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