Information on EC 2.4.99.19 - undecaprenyl-diphosphooligosaccharide-protein glycotransferase

Word Map on EC 2.4.99.19
Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
Specify your search results
Select one or more organisms in this record:
Show additional data
Do not include text mining results
Include (text mining) results (more...)
Include results (AMENDA + additional results, but less precise; more...)

The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
2.4.99.19
-
RECOMMENDED NAME
GeneOntology No.
undecaprenyl-diphosphooligosaccharide-protein glycotransferase
-
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
tritrans,heptacis-undecaprenyl diphosphooligosaccharide + [protein]-L-asparagine = tritrans,heptacis-undecaprenyl diphosphate + a glycoprotein with the oligosaccharide chain attached by N-beta-D-glycosyl linkage to protein L-asparagine
show the reaction diagram
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
protein N-glycosylation (bacterial)
-
-
SYSTEMATIC NAME
IUBMB Comments
tritrans,heptacis-undecaprenyl-diphosphooligosaccharide:protein-L-asparagine N-beta-D-oligosaccharidotransferase
A bacterial enzyme that has been isolated from Campylobacter jejuni [1] and Campylobacter lari [2]. It forms a glycoprotein by the transfer of a glucosyl-N-acetylgalactosaminyl-N,N'-diacetylbacillosamine (GalNAc2(Glc)GalNAc3diNAcBac) polysaccharide and related oligosaccharides to the side-chain of an L-asparagine residue in the sequence -Asp/Glu-Xaa-Asn-Xaa'-Ser/Thr- (Xaa and Xaa' not Pro) in nascent polypeptide chains. Requires Mn2+ or Mg2+. Occurs on the external face of the plasma membrane. The polyprenol involved is normally tritrans,heptacis-undecaprenol but a decaprenol is used by some species.
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
single gene pglB
-
-
Manually annotated by BRENDA team
gene pglB
UniProt
Manually annotated by BRENDA team
the organism contains two unrelated pglB genes, pglB1 and pglB2, neither of which is located within a larger locus involved in protein glycosylation
-
-
Manually annotated by BRENDA team
the organism contains two unrelated pglB genes, pglB1 and pglB2, neither of which is located within a larger locus involved in protein glycosylation
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
malfunction
metabolism
physiological function
additional information
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
N-acetyl-D-galactosaminyl-alpha-(1->3)-N,N'-diacetyl-alpha-D-bacillosaminyl-diphospho-tritrans,heptacis-undecaprenol + KDFNVSKA
tritrans,heptacis-undecaprenyl diphosphate + Lys-Asp-Phe-N4-[N-acetyl-D-galactosaminyl-alpha-(1->3)-N,N'-diacetyl-beta-D-bacillosaminyl]-Asn-Val-Ser-Lys-Ala
show the reaction diagram
-
oligosaccharyl transferase activity of PglB in vitro with synthetic disaccharide glycan donor and peptide acceptor substrate
-
-
?
tritrans,heptacis-undecaprenyl diphospho-O1 antigen + [protein]-L-asparagine
tritrans,heptacis-undecaprenyl diphosphate + [protein]-L-asparagine-O1 antigen
show the reaction diagram
-
from Shigella dysenteriae
a glycoprotein with the oligosaccharide chain attached by N-beta-D-glycosyl linkage to protein L-asparagine
-
?
tritrans,heptacis-undecaprenyl diphospho-O11 antigen + [protein]-L-asparagine
tritrans,heptacis-undecaprenyl diphosphate + [protein]-L-asparagine-O11 antigen
show the reaction diagram
-
from Pseudomonas aeruginosa
a glycoprotein with the oligosaccharide chain attached by N-beta-D-glycosyl linkage to protein L-asparagine
-
?
tritrans,heptacis-undecaprenyl diphospho-O157 antigen + [protein]-L-asparagine
tritrans,heptacis-undecaprenyl diphosphate + [protein]-L-asparagine-O157 antigen
show the reaction diagram
-
from Escherichia coli
a glycoprotein with the oligosaccharide chain attached by N-beta-D-glycosyl linkage to protein L-asparagine
-
?
tritrans,heptacis-undecaprenyl diphospho-O16 antigen + [protein]-L-asparagine
tritrans,heptacis-undecaprenyl diphosphate + [protein]-L-asparagine-O16 antigen
show the reaction diagram
-
from Escherichia coli
a glycoprotein with the oligosaccharide chain attached by N-beta-D-glycosyl linkage to protein L-asparagine
-
?
tritrans,heptacis-undecaprenyl diphospho-O7 antigen + [protein]-L-asparagine
tritrans,heptacis-undecaprenyl diphosphate + [protein]-L-asparagine-O7 antigen
show the reaction diagram
-
from Escherichia coli
a glycoprotein with the oligosaccharide chain attached by N-beta-D-glycosyl linkage to protein L-asparagine
-
?
tritrans,heptacis-undecaprenyl diphospho-O9 antigen + [Acra]-L-asparagine
tritrans,heptacis-undecaprenyl diphosphate + [Acra]-L-asparagine-O9 antigen
show the reaction diagram
-
PglB transfers the O9a antigen, an ABC transporter-dependent O antigen, to AcrA in Escherichia coli strain CWG28, the K30 antigen is not transferred to AcrA
a glycoprotein with the oligosaccharide chain attached by N-beta-D-glycosyl linkage to protein L-asparagine
-
?
tritrans,heptacis-undecaprenyl diphosphooligosaccharide + [HmcA]-L-asparagine
tritrans,heptacis-undecaprenyl diphosphate + [HmcA]-L-asparagine-oligosaccharide
show the reaction diagram
-
a protein from Desulfovibrio gigas, N-glycosylation site is N261
a glycoprotein with the oligosaccharide chain attached by N-beta-D-glycosyl linkage to protein L-asparagine
-
?
tritrans,heptacis-undecaprenyl diphosphooligosaccharide + [protein]-L-asparagine
tritrans,heptacis-undecaprenyl diphosphate + [protein]-L-asparagine-oligosaccharide
show the reaction diagram
[KDFNVSKA]-L-asparagine +N-acetyl-alpha-D-galactosaminyl-diphospho-tritrans,heptacis-undecaprenol
tritrans,heptacis-undecaprenyl diphosphate + Lys-Asp-Phe-N4-[N-acetyl-beta-D-galactosaminyl]-Asn-Val-Ser-Lys-Ala
show the reaction diagram
-
a chemically synthesized sugar donor with synthesized peptide acceptor
-
-
?
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
tritrans,heptacis-undecaprenyl diphosphooligosaccharide + [protein]-L-asparagine
tritrans,heptacis-undecaprenyl diphosphate + [protein]-L-asparagine-oligosaccharide
show the reaction diagram
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
-
x * 82000, SDS-PAGE
monomer
additional information
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
glycoprotein
functional PglB is partially autoglycosylated at N535 and N556
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
PglB crystal structure analysis
-
purified detagged and methylated wild-type and selenomethionine-labeled C-terminal globular domain of PglB, hanging drop vapour diffusion method, 500 nl of 10 mg/ml protein in 10 mM Tris-HCl, pH 8.0, is mixed in an 1:1 ratio with reservoir solution containing 0.1 M sodium cacodylate, pH 6.5, 18% PEG 8000, 0.2 M calcium acetate, 20°C, X-ray diffraction structure determination and analysis at 2.8 A resolution
PglB in complex with acceptor hexapeptide DQNATF, X-ray diffraction structure determination and analysis at 3.4 A resolution, molecular replacement using the periplasmic domain of Campylobacter jejuni PglB, PDB ID 3AAG, as model
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
recombinant His-tagged full-length PglB from Escherichia coli strain BL21(DE3) in the membrane fraction by ultracentrifugation, recombinant GST-tagged wild-type and selenomethionine-labeled C-terminal globular domain of PglB from Escherichia coli strain BL21(DE3)pLysS by glutathione affinity chromatography and cleavage of the tag by 3C protease, followed by to reductive methylation of the lysine residues, gel filtration, and anion exchange chromatography
recombinant His-tagged PglB from Escherichia coli strain C43(DE3) by nickel affinity chromatography
-
recombinant His10-tagged wild-type and mutant enzymes from Escherichia coli strain SCM6
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
gene pglB, expressio of the enzyme and the modified substrates in Escherichia coli
gene pglB, expression of His10-tagged wild-type and mutant enzymes in Escherichia coli strain SCM6
gene pglB, expression of the GST-tagged C-terminal globular domain of PglB in Escherichia coli strain BL21(DE3)pLysS, expression of His-tagged full-length PglB in Escherichia coli strain BL21(DE3)
gene pglB, expression of the wild-type and mutant WAAYGY-PglB variant in Escherichia coli in membranes
-
gene pglB, functional expression in Escherichia coli
-
gene pglB, functional expression in Escherichia coli strains E69, CWG28, or CWG44 using an arabinose-inducible promoter, coexpression of the Acra protein substrate
-
gene pglB, large-scale overexpression of His-tagged PglB in Escherichia coli strain C43(DE3)
-
gene pglB, sequence and structure comparison with STT3 subunit of the eukaryotic OTase complex of Saccharomyces cerevisiae
-
gene pglB, sequence comparisons and phylogenetic analysis, cloning and analysis of an operon that might encode all enzymes required for N-glycosylation in Desulfovibrio desulfuricans, functional expression of gene pglB in Eschericia coli
-
gene pglB, sequence comparisons and phylogenetic analysis, functional expression in Escherichia coli. All genes necessary for N-linked glycosylation in Campylobacter jejuni are located in a single locus
-
genes pglB1 and pglB2, DNA and amino acid sequence determination and analysis. Gene pglB1, but not the pglB2, gene is able to partially complement the Campylobacter jejuni pglB gene
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D152E
-
site-directed mutagenesis, D152 and E316 can both be mutated to their acidic counterparts (D152E and E316D) and still retain activity, albeit at a notably decreased level
D475A
-
site-directed mutagenesis, the mutant shows decreased activity compared to the wild-type enzyme
D475E
-
site-directed mutagenesis, the mutant shows decreased activity compared to the wild-type enzyme
D54A
-
site-directed mutagenesis, the increased Und-PP-Bac-GalNAc substrate concentration has little effect on the mutant activity
E316D
-
site-directed mutagenesis, D152 and E316 can both be mutated to their acidic counterparts (D152E and E316D) and still retain activity, albeit at a notably decreased level
E316Q
-
site-directed mutagenesis, inactive mutant
K478A
-
site-directed mutagenesis, the increased Und-PP-Bac-GalNAc substrate concentration has little effect on the mutant activity; site-directed mutagenesis, the mutant shows decreased activity compared to the wild-type enzyme
D154A
the mutation reduces the observed glycosylation yield by over 50% compared to the wild-type enzyme
D56A
the mutation reduces the observed glycosylation yield by over 90% compared to the wild-type enzyme
D56A/E319A
the double mutant is inactive
E319A
the mutation reduces the observed glycosylation yield by over 90% compared to the wild-type enzyme
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
medicine
-
application for protein glycosylation in recombinant bacteria in the production of potent conjugate vaccines where polysaccharide antigens of pathogenic bacteria are covalently bound to immunogenic carrier proteins