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Literature summary for 2.4.1.292 extracted from
- 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.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| 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
| Protein Variants | Comment | Organism |
|---|---|---|
| D170A | site-directed mutagenesis | Campylobacter jejuni |
| D306A | site-directed mutagenesis | Campylobacter jejuni |
| D307A | site-directed mutagenesis | Campylobacter jejuni |
| E171A | site-directed mutagenesis | Campylobacter jejuni |
| E179A | site-directed mutagenesis | Campylobacter jejuni |
| E265A | site-directed mutagenesis | Campylobacter jejuni |
| E273A | site-directed mutagenesis | Campylobacter jejuni |
| E308A | site-directed mutagenesis | Campylobacter jejuni |
| E316A | site-directed mutagenesis | Campylobacter jejuni |
| E346A | site-directed mutagenesis | Campylobacter jejuni |
| E354A | site-directed mutagenesis | Campylobacter jejuni |
| E41A | site-directed mutagenesis | Campylobacter jejuni |
| E49A | site-directed mutagenesis | Campylobacter jejuni |
| R191A | site-directed mutagenesis | Campylobacter jejuni |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 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
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Campylobacter jejuni | - |
gene pglH | - |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| 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)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 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
| Synonyms | Comment | Organism |
|---|---|---|
| PglH | - |
Campylobacter jejuni |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 8.5 | - |
assay at | Campylobacter jejuni |
General Information
| General Information | Comment | Organism |
|---|---|---|
| evolution | PglH is a glycosyltransferase protein containing the conserved EX7E sequence motif | Campylobacter jejuni |
| 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 |
| 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 |
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