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Literature summary extracted from

  • 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 (2009), Biochemistry, 48, 2807-2816.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.4.1.292 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 Campylobacter jejuni

Protein Variants

EC Number Protein Variants Comment Organism
2.4.1.292 D170A site-directed mutagenesis Campylobacter jejuni
2.4.1.292 D306A site-directed mutagenesis Campylobacter jejuni
2.4.1.292 D307A site-directed mutagenesis Campylobacter jejuni
2.4.1.292 E171A site-directed mutagenesis Campylobacter jejuni
2.4.1.292 E179A site-directed mutagenesis Campylobacter jejuni
2.4.1.292 E265A site-directed mutagenesis Campylobacter jejuni
2.4.1.292 E273A site-directed mutagenesis Campylobacter jejuni
2.4.1.292 E308A site-directed mutagenesis Campylobacter jejuni
2.4.1.292 E316A site-directed mutagenesis Campylobacter jejuni
2.4.1.292 E346A site-directed mutagenesis Campylobacter jejuni
2.4.1.292 E354A site-directed mutagenesis Campylobacter jejuni
2.4.1.292 E41A site-directed mutagenesis Campylobacter jejuni
2.4.1.292 E49A site-directed mutagenesis Campylobacter jejuni
2.4.1.292 R191A site-directed mutagenesis Campylobacter jejuni

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.4.1.292 3 UDP-N-acetyl-alpha-D-galactosamine + GalNAc-alpha-(1->4)-GalNAc-alpha-(1->3)-Bac2,4diNAc-diphospho-tritrans,heptacis-undecaprenol Campylobacter jejuni
-
 3 UDP + [GalNAc-alpha-(1->4)]4-GalNAc-alpha-(1->3)-Bac2,4diNAc-diphospho-tritrans,heptacis-undecaprenol
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.4.1.292 Campylobacter jejuni
-
 gene pglH
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.4.1.292 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 Campylobacter jejuni

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.4.1.292 3 UDP-N-acetyl-alpha-D-galactosamine + GalNAc-alpha-(1->4)-GalNAc-alpha-(1->3)-Bac2,4diNAc-diphospho-tritrans,heptacis-undecaprenol
-
 Campylobacter jejuni 3 UDP + [GalNAc-alpha-(1->4)]4-GalNAc-alpha-(1->3)-Bac2,4diNAc-diphospho-tritrans,heptacis-undecaprenol
-
 ?

Synonyms

EC Number Synonyms Comment Organism
2.4.1.292 PglH
-
 Campylobacter jejuni

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.4.1.292 8.5
-
 assay at Campylobacter jejuni

General Information

EC Number General Information Comment Organism
2.4.1.292 evolution PglH is a glycosyltransferase protein containing the conserved EX7E sequence motif Campylobacter jejuni
2.4.1.292 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 Campylobacter jejuni
2.4.1.292 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 Campylobacter jejuni