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Literature summary extracted from

  • Xu, C.; Liu, B.; Hu, B.; Han, Y.; Feng, L.; Allingham, J.S.; Szarek, W.A.
    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.
    View publication on PubMedView publication on EuropePMC

Activating Compound

EC Number Activating Compound Comment Organism Structure
2.4.1.304 2-mercaptoethanol 1-30 mM, 4fold-stimulation Shigella boydii
2.4.1.304 dithiothreitol 0.1-3 mM, about 5fold stimulation Shigella boydii

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.4.1.304 expressed in Escherichia coli Shigella boydii

Protein Variants

EC Number Protein Variants Comment Organism
2.4.1.304 D99A highly active mutant enzyme Shigella boydii
2.4.1.304 E101A mutant exhibits minimal enzyme activity, greatly reduced enzyme activity in the presence of either Mn2+ or Pb2+ Shigella boydii
2.4.1.304 E103A highly active mutant enzyme Shigella boydii
2.4.1.304 K211A greatly reduced activity, 60fold higher activity in the presence of Pb2+ than in the presence of Mn2+ Shigella boydii
2.4.1.304 R210A active mutant enzyme Shigella boydii
2.4.1.304 R212A active mutant enzyme Shigella boydii

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.4.1.304 4-hydroxyphenylglyoxal 0.2 mM, 50% inhibition Shigella boydii
2.4.1.304 5,5'-dithio-bis-2-nitrobenzoic acid 0.2 mM, 85% inhibition Shigella boydii
2.4.1.304 diethyl dicarbonate 0.2 mM 31% inhibition Shigella boydii
2.4.1.304 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]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.4.1.304 0.1
-
N-acetyl-alpha-D-glucosaminyl-diphospho-phenoxyundecanol pH 7, 37°C Shigella boydii
2.4.1.304 0.25
-
UDP-alpha-D-galactose pH 7, 37°C Shigella boydii

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.4.1.304 Ca2+ 5 mM, highest stimulation of activity in the presence of Mn2+. Ca2+, Pb2+, Ni2+ and Mg2+ are also effective Shigella boydii
2.4.1.304 Mg2+ 5 mM, highest stimulation of activityN in the presence of Mn2+. Ca2+, Pb2+, Ni2+ and Mg2+ are also effective Shigella boydii
2.4.1.304 Mn2+ 5 mM, highest stimulation of activity in the presence of Mn2+. Ca2+, Pb2+, Ni2+ and Mg2+ are also effective Shigella boydii
2.4.1.304 Ni2+ 5 mM, highest stimulation of activity in the presence of Mn2+. Ca2+, Pb2+, Ni2+ and Mg2+ are also effective Shigella boydii
2.4.1.304 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]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.4.1.304 30000
-
SDS-PAGE Shigella boydii

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.4.1.304 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
-
?
2.4.1.304 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

EC Number Organism UniProt Comment Textmining
2.4.1.304 Shigella boydii
-
-
-
2.4.1.304 Shigella boydii B14
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.4.1.304
-
Shigella boydii

Storage Stability

EC Number Storage Stability Organism
2.4.1.304 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)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.4.1.304 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 ?
-
?
2.4.1.304 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 ?
-
?
2.4.1.304 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
-
?
2.4.1.304 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
-
?
2.4.1.304 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
-
?
2.4.1.304 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
-
?
2.4.1.304 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
-
?
2.4.1.304 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
-
?
2.4.1.304 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
-
?
2.4.1.304 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
-
?
2.4.1.304 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
-
?
2.4.1.304 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

EC Number Subunits Comment Organism
2.4.1.304 ? x * 30000, SDS-PAGE, calculated from sequence Shigella boydii

Synonyms

EC Number Synonyms Comment Organism
2.4.1.304 UDP-Gal:GlcNAc-pyrophosphate-lipid beta-1,4-galactosyltransferase
-
Shigella boydii
2.4.1.304 UDP-Gal:GlcNAc-R 1,4-Gal-transferase
-
Shigella boydii
2.4.1.304 WfeD
-
Shigella boydii

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.4.1.304 37
-
assay at Shigella boydii

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.4.1.304 7
-
assay at Shigella boydii