Any feedback?
Enzyme Nomenclature
Information on EC 2.4.1.292 - GalNAc-alpha-(1->4)-GalNAc-alpha-(1->3)-diNAcBac-PP-undecaprenol alpha-1,4-N-acetyl-D-galactosaminyltransferase
for references in articles please use BRENDA:EC2.4.1.292Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
EC Tree
2.4.1.292 GalNAc-alpha-(1->4)-GalNAc-alpha-(1->3)-diNAcBac-PP-undecaprenol alpha-1,4-N-acetyl-D-galactosaminyltransferaseIUBMB Comments
Isolated from Campylobacter jejuni. Part of a bacterial N-linked glycosylation pathway.
Specify your search results
The enzyme appears in viruses and cellular organisms
Reaction Schemes
topPlease wait a moment until the data is sorted. This message will disappear when the data is sorted.
SYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
PglH
PglH
-
-
-
-
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
3 UDP-N-acetyl-alpha-D-galactosamine + GalNAc-alpha-(1->4)-GalNAc-alpha-(1->3)-diNAcBac-PP-tritrans,heptacis-undecaprenol = 3 UDP + [GalNAc-alpha-(1->4)]4-GalNAc-alpha-(1->3)-diNAcBac-PP-tritrans,heptacis-undecaprenol
3 UDP-N-acetyl-alpha-D-galactosamine + GalNAc-alpha-(1->4)-GalNAc-alpha-(1->3)-diNAcBac-PP-tritrans,heptacis-undecaprenol = 3 UDP + [GalNAc-alpha-(1->4)]4-GalNAc-alpha-(1->3)-diNAcBac-PP-tritrans,heptacis-undecaprenol
-
-
-
-
3 UDP-N-acetyl-alpha-D-galactosamine + GalNAc-alpha-(1->4)-GalNAc-alpha-(1->3)-diNAcBac-PP-tritrans,heptacis-undecaprenol = 3 UDP + [GalNAc-alpha-(1->4)]4-GalNAc-alpha-(1->3)-diNAcBac-PP-tritrans,heptacis-undecaprenol
PglH might follow a SN1-like mechanism, where the nucleophilic attack from the acceptor substrate and the departure of the leaving UDP occur from the same side. This would require the leaving group to act as a catalytic base to activate the incoming nucleophile. Glycan counting mechanism during PglH reaction, overview
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
PATHWAY SOURCE
PATHWAYS
-
-
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
SYSTEMATIC NAME
IUBMB Comments
UDP-N-acetyl-alpha-D-galactosamine:GalNAc-alpha-(1->4)-GalNAc-alphja-(1->3)-diNAcBac-PP-tritrans,heptacis-undecaprenol 4-alpha-N-acetyl-D-galactosaminyltransferase
Isolated from Campylobacter jejuni. Part of a bacterial N-linked glycosylation pathway.
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
3 UDP-GalNAc + GalNAc-alpha1,4-GalNAc-alpha1,3-Bac-alpha1-PP-undecaprenyl
3 UDP + (GalNAc-alpha1,4)4-GalNAc-alpha1,3-Bac-alpha1-PP-undecaprenyl
PglH-substrate binding structure, overview
-
-
?
3 UDP-N-acetyl-alpha-D-galactosamine + GalNAc-alpha-(1->4)-GalNAc-alpha-(1->3)-Bac2,4diNAc-diphospho-tritrans,heptacis-undecaprenol
3 UDP + [GalNAc-alpha-(1->4)]4-GalNAc-alpha-(1->3)-Bac2,4diNAc-diphospho-tritrans,heptacis-undecaprenol
N,N'-diacetylbacillosamine + glyceramido-acetamido trideoxyhexose
?
-
-
-
-
?
3 UDP-N-acetyl-alpha-D-galactosamine + GalNAc-alpha-(1->4)-GalNAc-alpha-(1->3)-Bac2,4diNAc-diphospho-tritrans,heptacis-undecaprenol
3 UDP + [GalNAc-alpha-(1->4)]4-GalNAc-alpha-(1->3)-Bac2,4diNAc-diphospho-tritrans,heptacis-undecaprenol
-
-
-
-
?
3 UDP-N-acetyl-alpha-D-galactosamine + GalNAc-alpha-(1->4)-GalNAc-alpha-(1->3)-Bac2,4diNAc-diphospho-tritrans,heptacis-undecaprenol
3 UDP + [GalNAc-alpha-(1->4)]4-GalNAc-alpha-(1->3)-Bac2,4diNAc-diphospho-tritrans,heptacis-undecaprenol
-
-
-
-
?
3 UDP-N-acetyl-alpha-D-galactosamine + GalNAc-alpha-(1->4)-GalNAc-alpha-(1->3)-Bac2,4diNAc-diphospho-tritrans,heptacis-undecaprenol
3 UDP + [GalNAc-alpha-(1->4)]4-GalNAc-alpha-(1->3)-Bac2,4diNAc-diphospho-tritrans,heptacis-undecaprenol
-
-
-
-
?
additional information
?
-
-
in vitro transfer of N,N'-diacetylbacillosamine phosphate onto undecaprenyl-phosphate and the in vitro enzymatic synthesis of the heptasacccharide using a chemically synthesized undecaprenyl diphosphate-linked N,N'-diacetylbacillosamine. The reactions of PglC, PglA, PglH, PglJ and PglI are coupled in a single reaction vessel, NMR spectrometric product analysis
-
-
?
additional information
?
-
-
in vitro, trisaccharide is prepared by using PglA and PglJ and then reacts singly with PglH
-
-
?
additional information
?
-
-
in vitro, trisaccharide is prepared by using PglA and PglJ and then reacts singly with PglH
-
-
?
additional information
?
-
-
PglH2 variant acts on both diNAcBac and GATDH to generate GlcNAc-containing disaccharides
-
-
-
additional information
?
-
-
PglH (from ORF3) acts as a glucosyltransferase, which uses both UndPP-linked diNAcBac and glyceramido-acetamido trideoxyhexose, GATDH
-
-
?
additional information
?
-
-
PglH (from ORF3) acts as a glucosyltransferase, which uses both undecaprenyl-diphospho-linked Bac2,4diNAc and glyceramido-acetamido trideoxyhexose, GATDH, glycosyltransferase specificity of PglH, overview. Recombinantly expressed PglH from strain Z2491 specifically transfers UDP-Glc to undecaprenyldiphospho-Bac2,4diNAc. PglH shows no transferase activity in the presence of UDP-Gal, UDP-GlcNAc, UDPGalNAc, and GDP-Man
-
-
?
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
3 UDP-N-acetyl-alpha-D-galactosamine + GalNAc-alpha-(1->4)-GalNAc-alpha-(1->3)-Bac2,4diNAc-diphospho-tritrans,heptacis-undecaprenol
3 UDP + [GalNAc-alpha-(1->4)]4-GalNAc-alpha-(1->3)-Bac2,4diNAc-diphospho-tritrans,heptacis-undecaprenol
3 UDP-N-acetyl-alpha-D-galactosamine + GalNAc-alpha-(1->4)-GalNAc-alpha-(1->3)-Bac2,4diNAc-diphospho-tritrans,heptacis-undecaprenol
3 UDP + [GalNAc-alpha-(1->4)]4-GalNAc-alpha-(1->3)-Bac2,4diNAc-diphospho-tritrans,heptacis-undecaprenol
-
-
-
-
?
3 UDP-N-acetyl-alpha-D-galactosamine + GalNAc-alpha-(1->4)-GalNAc-alpha-(1->3)-Bac2,4diNAc-diphospho-tritrans,heptacis-undecaprenol
3 UDP + [GalNAc-alpha-(1->4)]4-GalNAc-alpha-(1->3)-Bac2,4diNAc-diphospho-tritrans,heptacis-undecaprenol
-
-
-
-
?
3 UDP-N-acetyl-alpha-D-galactosamine + GalNAc-alpha-(1->4)-GalNAc-alpha-(1->3)-Bac2,4diNAc-diphospho-tritrans,heptacis-undecaprenol
3 UDP + [GalNAc-alpha-(1->4)]4-GalNAc-alpha-(1->3)-Bac2,4diNAc-diphospho-tritrans,heptacis-undecaprenol
-
-
-
-
?
additional information
?
-
-
PglH (from ORF3) acts as a glucosyltransferase, which uses both UndPP-linked diNAcBac and glyceramido-acetamido trideoxyhexose, GATDH
-
-
?
additional information
?
-
-
PglH (from ORF3) acts as a glucosyltransferase, which uses both undecaprenyl-diphospho-linked Bac2,4diNAc and glyceramido-acetamido trideoxyhexose, GATDH, glycosyltransferase specificity of PglH, overview. Recombinantly expressed PglH from strain Z2491 specifically transfers UDP-Glc to undecaprenyldiphospho-Bac2,4diNAc. PglH shows no transferase activity in the presence of UDP-Gal, UDP-GlcNAc, UDPGalNAc, and GDP-Man
-
-
?
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.002
(GalNAc-alpha1,4)4-GalNAc-alpha1,3-Bac-alpha1-PP-undecaprenyl
recombinant mutant K75A/R72A/K68A, pH 7.5, 25°C
0.006
(GalNAc-alpha1,4)4-GalNAc-alpha1,3-Bac-alpha1-PP-undecaprenyl
recombinant mutant E275A, pH 7.5, 25°C
0.008
(GalNAc-alpha1,4)4-GalNAc-alpha1,3-Bac-alpha1-PP-undecaprenyl
recombinant mutant E267A, pH 7.5, 25°C
0.039
(GalNAc-alpha1,4)4-GalNAc-alpha1,3-Bac-alpha1-PP-undecaprenyl
recombinant mutant K75A/R72A, pH 7.5, 25°C
0.15
(GalNAc-alpha1,4)4-GalNAc-alpha1,3-Bac-alpha1-PP-undecaprenyl
recombinant mutant R72A/K68A, pH 7.5, 25°C
0.19
(GalNAc-alpha1,4)4-GalNAc-alpha1,3-Bac-alpha1-PP-undecaprenyl
recombinant mutant H118A, pH 7.5, 25°C
0.19
(GalNAc-alpha1,4)4-GalNAc-alpha1,3-Bac-alpha1-PP-undecaprenyl
recombinant mutant K75A, pH 7.5, 25°C
0.31
(GalNAc-alpha1,4)4-GalNAc-alpha1,3-Bac-alpha1-PP-undecaprenyl
recombinant mutant R72A, pH 7.5, 25°C
0.34
(GalNAc-alpha1,4)4-GalNAc-alpha1,3-Bac-alpha1-PP-undecaprenyl
recombinant mutant T271A, pH 7.5, 25°C
0.69
(GalNAc-alpha1,4)4-GalNAc-alpha1,3-Bac-alpha1-PP-undecaprenyl
recombinant mutant K68A, pH 7.5, 25°C
2
(GalNAc-alpha1,4)4-GalNAc-alpha1,3-Bac-alpha1-PP-undecaprenyl
recombinant wild-type enzyme, pH 7.5, 25°C
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
and 11168H, hypermotile derivative of strains NCTC 11168 and 11828, gene pglH, cj1129
-
-
brenda
and 11168H, hypermotile derivative of strains NCTC 11168 and 11828, gene pglH, cj1129
-
-
brenda
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
evolution
malfunction
metabolism
physiological function
additional information
evolution
-
PglH is a glycosyltransferase protein containing the conserved EX7E sequence motif
evolution
-
genotyping, overview. The variable presence of two open reading frames (ORFs) in the pgl locus includes a putative glycosyltransferase gene, pglG, in addition to a glucosyltransferase-expressing gene, pglH. Polymorphisms exist at the gene level described for pglH and pglH2, where only one nonsynonymous mutation is accountable for the glycoform switch from Glc to GlcNAc. lacking these two ORFs retain the first 40 bp of pglG and the last 100 bp of pglH. Homologous recombination within the pgl loci, through genomic analysis of 100 African serogroup A isolates in Ethiopia in 2014, is detected representing the clonal replacement of hypervirulent meningococcal clone sequence type 7 (ST-7) by the ST-2859 descendant clone. Major polymorphism in pgl gene content within a small geographic area. This recombination event seems to emphasizes the role of protein glycosylation diversity in immune evasion. Polymorphisms exist at the gene level described for pglH and pglH2, where only one nonsynonymous mutation is accountable for the glycoform switch from Glc to GlcNAc. One subcluster has pglH while the other has the pglH2 variant allele
malfunction
-
pglH mutants have significantly reduced ability to adhere to and invade human epithelial Caco-2 cells, the 81116 pglH mutant is also severely affected in its ability to colonize chicks
malfunction
-
polymorphisms in PglH are associated with diminished glycosyltransferase activity
malfunction
-
pgl genotypes and glycosylation phenotypes in meningococcal isolates and the changes occurring during short-term asymptomatic carriage, overview. Polymorphisms exist at the gene level described for pglH and pglH2, where only one nonsynonymous mutation is accountable for the glycoform switch from Glc to GlcNAc
malfunction
-
pglH mutants have significantly reduced ability to adhere to and invade human epithelial Caco-2 cells, the 81116 pglH mutant is also severely affected in its ability to colonize chicks
-
metabolism
-
PglH is involved in the N-linked glycan structure formation, which is realized through a series of sequential glycosyl transfer reactions from uridine diphosphate-activated sugars to an undecaprenyl diphosphate carrier. Once undecaprenyl-diphospho-Bac2,4diNAc is formed by PglC, PglF, PglE, and PglD, sequential N-acetyl-galactosamine transfer reactions are catalyzed by PglA, PglJ and PglH to provide undecaprenyldiphospho-Bac2,4diNAc-GalNAc, undecaprenyldiphospho-Bac2,4diNAc-(GalNAc)2 and undecaprenyldiphospho-Bac2,4diNAc-(GalNAc)5, respectively
metabolism
-
PglH is part of a general N-linked glycosylation pathway, encoded by the pgl gene cluster, which culminates in the transfer of a heptasaccharide: GalNAc-alpha1,4-GalNAc-alpha1,4-(Glcbeta1,3)-GalNAc-alpha1,4-GalNAc-alpha1,4-GalNAc-alpha1,3-Bac, where Bac is bacillosamine (2,4-diacetamido-2,4,6-trideoxyglucose), from a membrane-anchored undecaprenylpyrophosphate-linked donor to the asparagine side chain of proteins at the Asn-X-Ser/Thr motif. The glycosyltransferases, PglA, PglH, PglI, and PglJ, responsible for the biosynthesis of the Und-PP-linked heptasaccharide. Elongation of the trisaccharide with PglH results in a hexasaccharide revealing the polymerase activity of PglH
metabolism
-
the Pgl pathway produces the N-linked glycan heptasaccharide GalNAc-alpha1,4-GalNAc-alpha1,4-(Glcbeta1,3)-GalNAc-alpha1,4-GalNAc-alpha1,4-GalNAc-alpha1,3-Bac2,4diNAc-1-Asn. The pathway begins with pyrophosphate bond formation between the Bac2,4diNAc phosphate and undecaprenylphosphate by Cj1124c (PglC) to form Bac2,4diNAc-alpha1-PP-Und. One N-acetylgalactosamine is linked to the polyisoprenediphosphate-bound N,N'-diacetylbacillosamine by Cj1125c (PglA) to form GalNAc-alpha1,3-Bac2,4diNAc-alpha1-PP-Und. Next, four additional GalNAc and one branching glucose are added sequentially to the isoprene-linked disaccharide by PglJ, PglH, and PglI, to form the heptasaccharide, overview
metabolism
the membrane-associated, processive and retaining glycosyltransferase PglH from Campylobacter jejuni is part of the biosynthetic pathway of the lipid-linked oligosaccharide (LLO) that serves as the glycan donor in bacterial protein N-glycosylation
metabolism
-
the Pgl pathway produces the N-linked glycan heptasaccharide GalNAc-alpha1,4-GalNAc-alpha1,4-(Glcbeta1,3)-GalNAc-alpha1,4-GalNAc-alpha1,4-GalNAc-alpha1,3-Bac2,4diNAc-1-Asn. The pathway begins with pyrophosphate bond formation between the Bac2,4diNAc phosphate and undecaprenylphosphate by Cj1124c (PglC) to form Bac2,4diNAc-alpha1-PP-Und. One N-acetylgalactosamine is linked to the polyisoprenediphosphate-bound N,N'-diacetylbacillosamine by Cj1125c (PglA) to form GalNAc-alpha1,3-Bac2,4diNAc-alpha1-PP-Und. Next, four additional GalNAc and one branching glucose are added sequentially to the isoprene-linked disaccharide by PglJ, PglH, and PglI, to form the heptasaccharide, overview
-
metabolism
-
PglH is part of a general N-linked glycosylation pathway, encoded by the pgl gene cluster, which culminates in the transfer of a heptasaccharide: GalNAc-alpha1,4-GalNAc-alpha1,4-(Glcbeta1,3)-GalNAc-alpha1,4-GalNAc-alpha1,4-GalNAc-alpha1,3-Bac, where Bac is bacillosamine (2,4-diacetamido-2,4,6-trideoxyglucose), from a membrane-anchored undecaprenylpyrophosphate-linked donor to the asparagine side chain of proteins at the Asn-X-Ser/Thr motif. The glycosyltransferases, PglA, PglH, PglI, and PglJ, responsible for the biosynthesis of the Und-PP-linked heptasaccharide. Elongation of the trisaccharide with PglH results in a hexasaccharide revealing the polymerase activity of PglH
-
physiological function
-
metabolic conflict related to competition for a shared substrate between the opposing glycosyltransferases PglA and PglH
physiological function
-
metabolic conflict related to competition for a shared substrate between the opposing glycosyltransferases PglA and PglH. Protein-linked PglH-derived disaccharides are immunogenic and antigenic, both diNAcBac- and GATDH-based PglH-derived glycans act as glycosyl donors in general, broad-spectrum protein glycosylation
physiological function
-
the glycosyl transfer polymerase is related to the virulence of Campylobacter jejuni, PglH is a single active site processive polymerase, that utilizes product inhibition to limit sequential glycosyl transfer reactions, the reactions cease after the addition of just three GalNAc residues, overview. Processive mechanism under initial rate conditions, but product inhibition and product accumulation led to PglH release of intermediate products prior to complete conversion to the native ultimate product. A single active site is responsible for all three transferase reactions, requirement of the EX7E motif in catalysis. Increased binding affinity with increasing glycan size is proposed to provide PglH with a counting mechanism that does not allow the transfer of more than three GalNAc residues, mechanismm overview
physiological function
a null mutation in pglH leads to the expression of a di-N-acetylbacillosamine-monosaccharide glycoform. PglH participates in the synthesis of undecaprenyldiphosphate-di-Nacetylbacillosamine-glucose
physiological function
PglH catalyzes the transfer of exactly three alpha1,4 N-acetylgalactosamine (Gal-NAc) units to the growing LLO precursor, GalNAc-alpha1,4-GalNAc-alpha1,3-Bac-alpha1-PP-undecaprenyl. PglH contains an amphipathic helix (ruler helix) that has a dual role of facilitating membrane attachment and glycan counting. The ruler helix contains three positively charged side chains that can bind the diphosphate group of the LLO substrate and thus limit the addition of GalNAc units to three, glycan counting mechanism during PglH reaction, overview
physiological function
-
PglH2 is a glucosyltransferase that acts on both N,N'-diacetylbacillosamine (diNAcBac) and glyceramido-acetamido trideoxyhexose (GATDH) to generate glucose GlcNAc-containing disaccharides
additional information
-
Pgl genotypes and phenotypes of strains, overview
additional information
-
Pgl genotypes and phenotypes of strains, overview. Epistatic interactions involving pglH at the intragenic and intergenic levels
additional information
-
architecture of PglH and donor substrate binding, molecular dynamics simulations, mechanism of glycan counting by PglH, overview. The structure of PglH bound to UDP-GalNAc reveals the molecular details of the interactions with the donor substrate. The uridine moiety is positioned in a pocket formed by alpha12 and the loop connecting alpha10 and alpha11. There are two hydrogen bonds of the uracil moiety with the main chain of V247. The 2' and 3' hydroxyl groups of the ribose moiety form hydrogen bonds with E275, which is part of the conserved EX7E motif along with E267, which interacts with the N-acetyl group of GalNAc. The diphosphate moiety interacts via hydrogen bonds with R191, K196, and T271. R191 and K196 are conserved in the GT-B family and are proposed to stabilize the leaving group (UDP) following the transfer reaction. The T271 side chain is involved in phosphate binding but does not have a catalytic role unlike R191 and K196
additional information
architecture of PglH and donor substrate binding, molecular dynamics simulations, mechanism of glycan counting by PglH, overview. The structure of PglH bound to UDP-GalNAc reveals the molecular details of the interactions with the donor substrate. The uridine moiety is positioned in a pocket formed by alpha12 and the loop connecting alpha10 and alpha11. There are two hydrogen bonds of the uracil moiety with the main chain of V247. The 2' and 3' hydroxyl groups of the ribose moiety form hydrogen bonds with E275, which is part of the conserved EX7E motif along with E267, which interacts with the N-acetyl group of GalNAc. The diphosphate moiety interacts via hydrogen bonds with R191, K196, and T271. R191 and K196 are conserved in the GT-B family and are proposed to stabilize the leaving group (UDP) following the transfer reaction. The T271 side chain is involved in phosphate binding but does not have a catalytic role unlike R191 and K196
additional information
-
more than 30 different glycoforms can be synthesized by combinations of glycosyltransferases and the O-acetylase within neisserial species
Show AA Sequence and Transmembrane Helices (402 entries)
Please use the AA Sequence and Transmembrane Helices Search for a specific query.
Please use the AA Sequence and Transmembrane Helices Search for a specific query.
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
additional information
-
PglH exhibits the protein fold described for glycosyltransferases GT-B GTs, consisting of two Rossman-like domains connected by a linker region. The conformation of PglH is similar in all three structures, although a slight rotation of the C-terminal vs. the N-terminal domain is observed in the ternary complex, where PglH is bound to UDP and LLO analogues
additional information
PglH exhibits the protein fold described for glycosyltransferases GT-B GTs, consisting of two Rossman-like domains connected by a linker region. The conformation of PglH is similar in all three structures, although a slight rotation of the C-terminal vs. the N-terminal domain is observed in the ternary complex, where PglH is bound to UDP and LLO analogues
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
crystal structures of PglH in a binary complex with UDP-GalNAc and two ternary complexes containing a chemo-enzymatically generated lipid-linked oligosaccharide analogue and either UDP or synthetic, nonhydrolyzable UDP-CH2-GalNAc. PglH contains an amphipathic helix that has a dual role of facilitating membrane attachment and glycan counting. The helix contains three positively charged side chains that can bind the diphosphate group of the lipid-linked oligosaccharide substrate and thus limit the addition of GalNAc units to three
purified recombinant His-tagged PglH (native or the selenomethionine derivative) in three distinct states, including a binary complex with UDP-GalNAc and two ternary complexes containing a chemo-enzymatically generated LLO analogue and either UDP or synthetic, nonhydrolyzable UDP-CH2-GalNAc, apoenzyme by vapor diffusion in sitting or hanging drops at 20°C, using a reservoir containing 0.2M KI, 0.1M MOPS, pH 7.2, and 17% PEG 3350, the volume ratio of protein to reservoir is 1:1, for PglH with bound acceptor substrate, concentrated PglH is incubated with synthetic LLO at 0.6 mM and UDP or UDP-CH2-GalNAc at 2 mM (final concentrations) for 15 min, crystallization is performed by vapor diffusion in sitting drops or hanging drops at 20°C using a reservoir containing 0.2 M Ammonium acetate, 0.1 M ADA 6.3, and 21-22% PEG 3350, the volume ratio of protein to reservoir is 1:1, 1-4 days, X-ray diffraction structure determination and analysis at 2.3-3.3 A resolution
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
E267A
site-directed mutagenesis, the mutant shows an about 300fold decrease in the turnover rate compared to the wild type enzyme
E275A
site-directed mutagenesis, the mutant shows an about 300fold decrease in the turnover rate compared to the wild type enzyme
H118A
site-directed mutagenesis, the mutant exhibits a reduced turnover rate compared to wild-type
K68A
site-directed mutagenesis, the mutant exhibits a reduced turnover rate compared to wild-type
K75A
site-directed mutagenesis, the mutant exhibits a reduced turnover rate compared to wild-type
K75A, R72A
site-directed mutagenesis, the mutant exhibits a reduced turnover rate compared to wild-type
K75A/R72A/K68A
site-directed mutagenesis, the mutant exhibits a reduced turnover rate compared to wild-type
R191A
R72A
site-directed mutagenesis, the mutant exhibits a reduced turnover rate compared to wild-type
R72A/K68A
site-directed mutagenesis, the mutant exhibits a reduced turnover rate compared to wild-type
T271A
site-directed mutagenesis, the mutant exhibits a fivefold reduction in turnover rate compared to wild-type
H371R
-
site-directed mutagenesis, strain FA 1090, the mutation is sufficient to restore PglH activity in the wild-type FA1090 strain
additional information
additional information
-
construction of pglH::kanr mutants, and complementation studies. The pglH::kanr mutant of strain 81116 has reduced colonization efficiency in a chick model. Mutation of pglH in strains 11168H and 81116 results in the reduced attachment to and invasion of Caco-2 cells. Reduction in attachment and invasion for strain 81116 are 23fold and 15fold, respectively, whereas the corresponding figures for strain 11168H are 6fold and and 5.4fold, respectively
additional information
-
construction of pglH::kanr mutants, and complementation studies. The pglH::kanr mutant of strain 81116 has reduced colonization efficiency in a chick model. Mutation of pglH in strains 11168H and 81116 results in the reduced attachment to and invasion of Caco-2 cells. Reduction in attachment and invasion for strain 81116 are 23fold and 15fold, respectively, whereas the corresponding figures for strain 11168H are 6fold and and 5.4fold, respectively
-
additional information
-
replacement of the strain N400 locus carrying the deleted form of ORFs 2 and 3 with those derived from the Neisseria gonorrhoeae strain FA1090 and from diverse neisserial strains into Neisseria gonorrhoeae strain N400 in which the pgl gene function and associated glycan structures is defined, mutant glycosylation patterns, overview. The presence of a stereotypic, conserved deletion mutation is inactivating pglH in strains of Neisseria gonorrhoeae, Neisseria meningitidis, and related commensals
additional information
-
replacement of the strain N400 locus carrying the deleted form of ORFs 2 and 3 with those derived from the Neisseria gonorrhoeae strain FA1090. The presence of a stereotypic, conserved deletion mutation is inactivating pglH in strains of Neisseria gonorrhoeae, Neisseria meningitidis, and related commensals
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
recombinant His-tagged PglH from Escherichia coli by nickel affinity chromatography
-
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21-Gold (DE3) by ultracentrifugation, nickel affinity chromatography, desalting gel filtration, gel filtration, and ultratfiltration
recombinant N-terminally octahistidine-tagged enzyme from Escherichia coli strain BL-21 RIL by nickel affinity chromatography, followed by cleavage of the His8-tag by TEV protease and another step of nickel affinity chromatography
-
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
gene pglH, expression of the N-terminally octahistidine-tagged enzyme containing a TEV cleavage sequence in Escherichia coli strain BL-21 RIL using a modified auto-induction method
-
gene pglH, recombinant expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21-Gold (DE3)
gene pglH2, genotyping, overview. The variable presence of two open reading frames (ORFs) in the pgl locus includes a putative glycosyltransferase gene, pglG, in addition to a glucosyltransferase-expressing gene, pglH. One subcluster has pglH while the other has the pglH2 variant allele. Strains lacking these two ORFs retain the first 40 bp of pglG and the last 100 bp of pglH. Homologous recombination within the pgl loci through genomic analysis of 100 African serogroup A isolates is detected representing the clonal replacement of hypervirulent meningococcal clone sequence type 7 (ST-7) by the ST-2859 descendant clone. Polymorphisms exist at the gene level described for pglH and pglH2, where only one nonsynonymous mutation is accountable for the glycoform switch from Glc to GlcNAc. One subcluster has pglH while the other has the pglH2 variant allele. The pglH locus shows polymorphisms between STs along the whole gene. A 5' point mutation (guanosine to cytosine) adjacent to the polycytosine [poly(C)] tract is found in individuals 20, 22, and 33 carrying ST-192 isolates
-
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Olivier, N.B.; Chen, M.M.; Behr, J.R.; Imperiali, B.
In vitro biosynthesis of UDP-N,N'-diacetylbacillosamine by enzymes of the Campylobacter jejuni general protein glycosylation system
Biochemistry
45
13659-13669
2006
Campylobacter jejuni, Campylobacter jejuni NCTC 11168
brenda
Troutman, J.M.; Imperiali, B.
Campylobacter jejuni PglH is a single active site processive polymerase that utilizes product inhibition to limit sequential glycosyl transfer reactions
Biochemistry
48
2807-2816
2009
Campylobacter jejuni
brenda
Karlyshev, A.V.; Everest, P.; Linton, D.; Cawthraw, S.; Newell, D.G.; Wren, B.W.
The Campylobacter jejuni general glycosylation system is important for attachment to human epithelial cells and in the colonization of chicks
Microbiology
150
1957-1964
2004
Campylobacter jejuni, Campylobacter jejuni NCTC 11168
brenda
Glover, K.J.; Weerapana, E.; Imperiali, B.
In vitro assembly of the undecaprenylpyrophosphate-linked heptasaccharide for prokaryotic N-linked glycosylation
Proc. Natl. Acad. Sci. USA
102
14255-14259
2005
Campylobacter jejuni, Campylobacter jejuni NCTC 11168
brenda
Boerud, B.; Viburiene, R.; Hartley, M.D.; Paulsen, B.S.; Egge-Jacobsen, W.; Imperiali, B.; Koomey, M.
Genetic and molecular analyses reveal an evolutionary trajectory for glycan synthesis in a bacterial protein glycosylation system
Proc. Natl. Acad. Sci. USA
108
9643-9648
2011
Neisseria gonorrhoeae, Neisseria meningitidis
brenda
Anonsen, J.; Vik, A.; Borud, B.; Viburiene, R.; Aas, F.; Kidd, S.; Aspholm, M.; Koomey, M.
Characterization of a unique tetrasaccharide and distinct glycoproteome in the O-linked protein glycosylation system of Neisseria elongata subsp. glycolytica
J. Bacteriol.
198
256-267
2016
Neisseria elongata subsp. glycolytica (A0A0N7IL44)
brenda
Borud, B.; Barnes, G.; Brynildsrud, O.; Fritzsonn, E.; Caugant, D.
Genotypic and phenotypic characterization of the O-linked protein glycosylation system reveals high glycan diversity in paired meningococcal carriage isolates
J. Bacteriol.
200
e00794-17
2018
Neisseria elongata subsp. glycolytica
brenda
Ramirez, A.; Boilevin, J.; Mehdipour, A.; Hummer, G.; Darbre, T.; Reymond, J.; Locher, K.
Structural basis of the molecular ruler mechanism of a bacterial glycosyltransferase
Nat. Commun.
9
445
2018
Campylobacter jejuni, Campylobacter jejuni (O86151)
brenda
Select items on the left to see more content.
EXTERNAL LINKS (specific for EC-Number 2.4.1.292)
html completed
Use of this online version of BRENDA is free under the CC BY 4.0 license. See terms of use for full details.
Release 2023.1 (January 2023)
About BRENDA
Social media
Help
Survey
Download
Contribution
Legal
Curated at
Member of
