Dataset Information

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.

SUBMITTER: Sun G

PROVIDER: S-EPMC4367255 | biostudies-literature | 2015 Mar

REPOSITORIES: biostudies-literature

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Identification and characterization of a novel prokaryotic peptide: N-glycosidase from Elizabethkingia meningoseptica.

Sun Guiqin G Yu Xiang X Bao Celimuge C Wang Lei L Li Meng M Gan Jianhua J Qu Di D Ma Jinbiao J Chen Li L

The Journal of biological chemistry 20150122 12

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 subj ...[more]

PMID: 25614628

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Project description:Streptococcus intermedius is a known human pathogen and belongs to the anginosus group (S. anginosus, S. intermedius, and S. constellatus) of streptococci (AGS). We found a large open reading frame (6,708 bp) in the lac operon, and bioinformatic analysis suggested that this gene encodes a novel glycosidase that can exhibit ?-d-galactosidase and N-acetyl-?-d-hexosaminidase activities. We, therefore, named this protein "multisubstrate glycosidase A" (MsgA). To test whether MsgA has these glycosidase activities, the msgA gene was disrupted in S. intermedius. The msgA-deficient mutant no longer showed cell- and supernatant-associated ?-d-galactosidase, ?-d-fucosidase, N-acetyl-?-d-glucosaminidase, and N-acetyl-?-d-galactosaminidase activities, and all phenotypes were complemented in trans with a recombinant plasmid carrying msgA. Purified MsgA had all four of these glycosidase activities and exhibited the lowest Km with 4-methylumbelliferyl-linked N-acetyl-?-d-glucosaminide and the highest kcat with 4-methylumbelliferyl-linked ?-d-galactopyranoside. In addition, the purified LacZ domain of MsgA had ?-d-galactosidase and ?-d-fucosidase activities, and the GH20 domain exhibited both N-acetyl-?-d-glucosaminidase and N-acetyl-?-d-galactosaminidase activities. The ?-d-galactosidase and ?-d-fucosidase activities of MsgA are thermolabile, and the optimal temperature of the reaction was 40°C, whereas almost all enzymatic activities disappeared at 49°C. The optimal temperatures for the N-acetyl-?-d-glucosaminidase and N-acetyl-?-d-galactosaminidase activities were 58 and 55°C, respectively. The requirement of sialidase treatment to remove sialic acid residues of the glycan branch end for glycan degradation by MsgA on human ?1-antitrypsin indicates that MsgA has exoglycosidase activities. MsgA and sialidase might have an important function in the production and utilization of monosaccharides from oligosaccharides, such as glycans for survival in a normal habitat and for pathogenicity of S. intermedius.

Dataset Information

Identification and characterization of a novel prokaryotic peptide: N-glycosidase from Elizabethkingia meningoseptica.

Publications

Identification and characterization of a novel prokaryotic peptide: N-glycosidase from Elizabethkingia meningoseptica.

Similar Datasets

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