title: Identification and characterization of a novel prokaryotic peptide: N-glycosidase from Elizabethkingia meningoseptica.
abstract: Peptide:N-glycosidase (PNGase) F, the first PNGase identified in prokaryotic cells, catalyzes the removal of intact asparagine-linked oligosaccharide chains from glycoproteins and/or glycopeptides. Since its discovery in 1984, PNGase F has remained as the sole prokaryotic PNGase. Recently, a novel gene encoding a protein with a predicted PNGase domain was identified from a clinical isolate of Elizabethkingia meningoseptica. In this study, the candidate protein was expressed in vitro and was subjected to biochemical and structural analyses. The results revealed that it possesses PNGase activity and has substrate specificity different from that of PNGase F. The crystal structure of the protein was determined at 1.9 Å resolution. Structural comparison with PNGase F revealed a relatively larger glycan-binding groove in the catalytic domain and an additional bowl-like domain with unknown function at the N terminus of the candidate protein. These structural and functional analyses indicated that the candidate protein is a novel prokaryotic N-glycosidase. The protein has been named PNGase F-II.
authors.affiliation: From the Key Laboratory of Medical Molecular Virology, Ministry of Education and Health, Shanghai Medical College, Fudan University, Shanghai 200032, China, Zhejiang Chinese Medical University, Hangzhou 310053, China.
authors.forename: Guiqin
authors.initials: G
authors.lastname: Sun
authors.affiliation: the State Key Laboratory of Genetic Engineering, Collaborative Innovation Center of Genetics and Development, School of Life Sciences, Fudan University, Shanghai 200438, China.
authors.forename: Xiang
authors.initials: X
authors.lastname: Yu
authors.affiliation: From the Key Laboratory of Medical Molecular Virology, Ministry of Education and Health, Shanghai Medical College, Fudan University, Shanghai 200032, China.
authors.forename: Celimuge
authors.initials: C
authors.lastname: Bao
authors.affiliation: From the Key Laboratory of Medical Molecular Virology, Ministry of Education and Health, Shanghai Medical College, Fudan University, Shanghai 200032, China.
authors.forename: Lei
authors.initials: L
authors.lastname: Wang
authors.affiliation: From the Key Laboratory of Medical Molecular Virology, Ministry of Education and Health, Shanghai Medical College, Fudan University, Shanghai 200032, China.
authors.forename: Meng
authors.initials: M
authors.lastname: Li
authors.affiliation: the Department of Physiology and Biophysics, School of Life Sciences, Fudan University, Shanghai 200433, China, and.
authors.forename: Jianhua
authors.initials: J
authors.lastname: Gan
authors.affiliation: From the Key Laboratory of Medical Molecular Virology, Ministry of Education and Health, Shanghai Medical College, Fudan University, Shanghai 200032, China.
authors.forename: Di
authors.initials: D
authors.lastname: Qu
authors.affiliation: the State Key Laboratory of Genetic Engineering, Collaborative Innovation Center of Genetics and Development, School of Life Sciences, Fudan University, Shanghai 200438, China, majb@fudan.edu.cn.
authors.forename: Jinbiao
authors.initials: J
authors.lastname: Ma
authors.affiliation: From the Key Laboratory of Medical Molecular Virology, Ministry of Education and Health, Shanghai Medical College, Fudan University, Shanghai 200032, China, lichen_bk@fudan.edu.cn.
authors.forename: Li
authors.initials: L
authors.lastname: Chen
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D018360:Crystallography, X-Ray; D041965:Flavobacteriaceae / Q000201:enzymology*; D006031:Glycosylation; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase / Q000737:chemistry / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity
chemical_list: D009838:Oligodeoxyribonucleotides; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase
keywords: Bacteria; Crystal Structure; Enzyme; Glycoprotein; N-Linked Glycosylation; Peptide:N-Glycosidase (PNGase), PNGase F, Glycosylation, Deglycosylation, Glycoprotein, Crystal Structure