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Literature summary for 2.4.1.304 extracted from
- Wang, L.; Brockhausen, I.: Biochemical characterization of UDP-Gal:GlcNAc-pyrophosphate-lipid beta-1,4-Galactosyltransferase WfeD, a new enzyme from Shigella boydii type 14 that catalyzes the second step in O-antigen repeating-unit synthesis (2010), J. Bacteriol., 193, 449-459.
Activating Compound
| Activating Compound | Comment | Organism | Structure |
|---|---|---|---|
| 2-mercaptoethanol | 1-30 mM, 4fold-stimulation | Shigella boydii |  |
| dithiothreitol | 0.1-3 mM, about 5fold stimulation | Shigella boydii | |
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| expressed in Escherichia coli | Shigella boydii |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| D99A | highly active mutant enzyme | Shigella boydii |
| E101A | mutant exhibits minimal enzyme activity, greatly reduced enzyme activity in the presence of either Mn2+ or Pb2+ | Shigella boydii |
| E103A | highly active mutant enzyme | Shigella boydii |
| K211A | greatly reduced activity, 60fold higher activity in the presence of Pb2+ than in the presence of Mn2+ | Shigella boydii |
| R210A | active mutant enzyme | Shigella boydii |
| R212A | active mutant enzyme | Shigella boydii |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| 4-hydroxyphenylglyoxal | 0.2 mM, 50% inhibition | Shigella boydii | |
| 5,5'-dithio-bis-2-nitrobenzoic acid | 0.2 mM, 85% inhibition | Shigella boydii | |
| diethyl dicarbonate | 0.2 mM 31% inhibition | Shigella boydii | |
| Triton X-100 | as the concentration of Triton X-100 increases from 0 to 0.5% in the reaction mixture, the activity of the enzyme decreases but is maintained at about 60% of the original activity, up to 6% Triton X-100 | Shigella boydii | |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.1 | - |
N-acetyl-alpha-D-glucosaminyl-diphospho-phenoxyundecanol | pH 7, 37°C | Shigella boydii | |
| 0.25 | - |
UDP-alpha-D-galactose | pH 7, 37°C | Shigella boydii | |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Ca2+ | 5 mM, highest stimulation of activity in the presence of Mn2+. Ca2+, Pb2+, Ni2+ and Mg2+ are also effective | Shigella boydii | |
| Mg2+ | 5 mM, highest stimulation of activityN in the presence of Mn2+. Ca2+, Pb2+, Ni2+ and Mg2+ are also effective | Shigella boydii | |
| Mn2+ | 5 mM, highest stimulation of activity in the presence of Mn2+. Ca2+, Pb2+, Ni2+ and Mg2+ are also effective | Shigella boydii | |
| Ni2+ | 5 mM, highest stimulation of activity in the presence of Mn2+. Ca2+, Pb2+, Ni2+ and Mg2+ are also effective | Shigella boydii | |
| Pb2+ | 5 mM, highest stimulation of activity in the presence of Mn2+. Ca2+, Pb2+, Ni2+ and Mg2+ are also effective | Shigella boydii | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 30000 | - |
SDS-PAGE | Shigella boydii |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| UDP-alpha-D-galactose + N-acetyl-alpha-D-glucosaminyl-diphospho-ditrans,octacis-undecaprenol | Shigella boydii | the O-antigen in the outer membrane of Gram-negative bacteria contains many repeats of an oligosaccharide unit. WfeD catalyzes a step of the repeating-unit assembly in Shigella boydii B14 | UDP + beta-D-Gal-(1->4)-alpha-D-GlcNAc-diphospho-ditrans,octacis-undecaprenol | - |
? | |
| UDP-alpha-D-galactose + N-acetyl-alpha-D-glucosaminyl-diphospho-ditrans,octacis-undecaprenol | Shigella boydii B14 | the O-antigen in the outer membrane of Gram-negative bacteria contains many repeats of an oligosaccharide unit. WfeD catalyzes a step of the repeating-unit assembly in Shigella boydii B14 | UDP + beta-D-Gal-(1->4)-alpha-D-GlcNAc-diphospho-ditrans,octacis-undecaprenol | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Shigella boydii | - |
- |
- |
| Shigella boydii B14 | - |
- |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
- |
Shigella boydii |
Storage Stability
| Storage Stability | Organism |
|---|---|
| 4°C, the crude enzyme preparation retains at least 60% of the initial activity after storage for up to 9 weeks | Shigella boydii |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| additional information | no activity with: alpha- and beta-anomers of GlcNAc-benzyl, beta-anomers of GlcNBu-benzyl, GlcNAc-naphthyl, GlcNAc-isoquinolinyl, as well as GlcNAc-alpha-OCOCH2-COO(CH2)11-OPh and alpha- and beta-anomers of GlcNAc-(CH2)11-OPh | Shigella boydii | ? | - |
? | |
| additional information | no activity with: alpha- and beta-anomers of GlcNAc-benzyl, beta-anomers of GlcNBu-benzyl, GlcNAc-naphthyl, GlcNAc-isoquinolinyl, as well as GlcNAc-alpha-OCOCH2-COO(CH2)11-OPh and alpha- and beta-anomers of GlcNAc-(CH2)11-OPh | Shigella boydii B14 | ? | - |
? | |
| UDP-alpha-D-galactose + N-acetyl-alpha-D-glucosaminyl-diphospho-ditrans,octacis-undecaprenol | the O-antigen in the outer membrane of Gram-negative bacteria contains many repeats of an oligosaccharide unit. WfeD catalyzes a step of the repeating-unit assembly in Shigella boydii B14 | Shigella boydii | UDP + beta-D-Gal-(1->4)-alpha-D-GlcNAc-diphospho-ditrans,octacis-undecaprenol | - |
? | |
| UDP-alpha-D-galactose + N-acetyl-alpha-D-glucosaminyl-diphospho-ditrans,octacis-undecaprenol | the O-antigen in the outer membrane of Gram-negative bacteria contains many repeats of an oligosaccharide unit. WfeD catalyzes a step of the repeating-unit assembly in Shigella boydii B14 | Shigella boydii B14 | UDP + beta-D-Gal-(1->4)-alpha-D-GlcNAc-diphospho-ditrans,octacis-undecaprenol | - |
? | |
| UDP-alpha-D-galactose + N-acetyl-alpha-D-glucosaminyl-diphospho-hexadecanol | the enzyme requires diphosphate in the acceptor substrate | Shigella boydii | UDP + beta-D-Gal-(1->4)-N-acetyl-alpha-D-glucosaminyl-diphospho-hexadecanol | - |
? | |
| UDP-alpha-D-galactose + N-acetyl-alpha-D-glucosaminyl-diphospho-hexadecanol | the enzyme requires diphosphate in the acceptor substrate | Shigella boydii B14 | UDP + beta-D-Gal-(1->4)-N-acetyl-alpha-D-glucosaminyl-diphospho-hexadecanol | - |
? | |
| UDP-alpha-D-galactose + N-acetyl-alpha-D-glucosaminyl-diphospho-hexanol | the enzyme requires diphosphate in the acceptor substrate | Shigella boydii | UDP + beta-D-Gal-(1->4)-N-acetyl-alpha-D-glucosaminyl-diphospho-hexanol | - |
? | |
| UDP-alpha-D-galactose + N-acetyl-alpha-D-glucosaminyl-diphospho-hexanol | the enzyme requires diphosphate in the acceptor substrate | Shigella boydii B14 | UDP + beta-D-Gal-(1->4)-N-acetyl-alpha-D-glucosaminyl-diphospho-hexanol | - |
? | |
| UDP-alpha-D-galactose + N-acetyl-alpha-D-glucosaminyl-diphospho-phenoxyundecanol | the synthetic acceptor substrate D-GlcNAc-alpha-PP-(CH2)11-O-Ph is an amphipathic molecule with similarity to the endogenous substrate D-GlcNAc-alpha-diphospho-ditrans,octacis-undecaprenol. The enzyme requires diphosphate in the acceptor substrate, Strict specificity for UDP-D-galactose as donor substrate | Shigella boydii | UDP + beta-D-Gal-(1->4)-N-acetyl-alpha-D-glucosaminyl-diphospho-phenoxyundecanol | - |
? | |
| UDP-alpha-D-galactose + N-acetyl-alpha-D-glucosaminyl-diphosphononanol | the enzyme requires diphosphate in the acceptor substrate | Shigella boydii | UDP + beta-D-Gal-(1->4)-N-acetyl-alpha-D-glucosaminyl-diphosphononanol | - |
? | |
| UDP-alpha-D-galactose + N-acetyl-alpha-D-glucosaminyl-diphosphononanol | the enzyme requires diphosphate in the acceptor substrate | Shigella boydii B14 | UDP + beta-D-Gal-(1->4)-N-acetyl-alpha-D-glucosaminyl-diphosphononanol | - |
? | |
| UDP-alpha-D-galactose + N-acetyl-alpha-D-glucosaminyl-phospho-phenoxyundecanol | synthetic acceptor. 11% of the activity with N-acetyl-D-glucosaminyl-alpha-diphospho-phenoxyundecanol | Shigella boydii | UDP + beta-D-Gal-(1->4)-N-acetyl-alpha-D-glucosaminyl-phospho-phenoxyundecanol | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| ? | x * 30000, SDS-PAGE, calculated from sequence | Shigella boydii |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| UDP-Gal:GlcNAc-pyrophosphate-lipid beta-1,4-galactosyltransferase | - |
Shigella boydii |
| UDP-Gal:GlcNAc-R 1,4-Gal-transferase | - |
Shigella boydii |
| WfeD | - |
Shigella boydii |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 37 | - |
assay at | Shigella boydii |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7 | - |
assay at | Shigella boydii |
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