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Information on EC 2.6.1.98 - UDP-2-acetamido-2-deoxy-ribo-hexuluronate aminotransferase and Organism(s) Pseudomonas aeruginosa and UniProt Accession Q9HZ76

for references in articles please use BRENDA:EC2.6.1.98
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IUBMB Comments
A pyridoxal 5'-phosphate protein. This enzyme participates in the biosynthetic pathway for UDP-alpha-D-ManNAc3NAcA (UDP-2,3-diacetamido-2,3-dideoxy-alpha-D-mannuronic acid), an important precursor of B-band lipopolysaccharide. The enzymes from Pseudomonas aeruginosa serotype O5 and Thermus thermophilus form a complex with the previous enzyme in the pathway, EC 1.1.1.335 (UDP-N-acetyl-2-amino-2-deoxyglucuronate oxidase).
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Pseudomonas aeruginosa
UNIPROT: Q9HZ76
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The taxonomic range for the selected organisms is: Pseudomonas aeruginosa
The enzyme appears in selected viruses and cellular organisms
Synonyms
aminotransferase wbpe, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
aminotransferase WbpE
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SYSTEMATIC NAME
IUBMB Comments
UDP-2-acetamido-3-amino-2,3-dideoxy-alpha-D-glucuronate:2-oxoglutarate aminotransferase
A pyridoxal 5'-phosphate protein. This enzyme participates in the biosynthetic pathway for UDP-alpha-D-ManNAc3NAcA (UDP-2,3-diacetamido-2,3-dideoxy-alpha-D-mannuronic acid), an important precursor of B-band lipopolysaccharide. The enzymes from Pseudomonas aeruginosa serotype O5 and Thermus thermophilus form a complex with the previous enzyme in the pathway, EC 1.1.1.335 (UDP-N-acetyl-2-amino-2-deoxyglucuronate oxidase).
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
UDP-2-acetamido-3-amino-2,3-dideoxy-alpha-D-glucuronate + 2-oxoglutarate
UDP-2-acetamido-2-deoxy-alpha-D-ribo-hex-3-uluronate + L-glutamate
show the reaction diagram
UDP-2-acetamido-3-amino-2,3-dideoxy-alpha-D-glucuronate + 2-oxoglutarate
UDP-2-acetamido-2-deoxy-alpha-D-ribo-hex-3-uluronate + L-glutamate
show the reaction diagram
additional information
?
-
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
UDP-2-acetamido-3-amino-2,3-dideoxy-alpha-D-glucuronate + 2-oxoglutarate
UDP-2-acetamido-2-deoxy-alpha-D-ribo-hex-3-uluronate + L-glutamate
show the reaction diagram
UDP-2-acetamido-3-amino-2,3-dideoxy-alpha-D-glucuronate + 2-oxoglutarate
UDP-2-acetamido-2-deoxy-alpha-D-ribo-hex-3-uluronate + L-glutamate
show the reaction diagram
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
pyridoxal 5'-phosphate
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pyridoxal 5'-phosphate
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-
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7 - 8
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assay at
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
WbpE is a member of the broad class of fold type I aspartate-aminotransferase enzymes, which harness the powerful electron-sink properties of PLP to carry out a wide variety of transformations, including transaminations, eliminations, decarboxylations, and racemizations. The general mechanism of this class of enzymes has been worked out in great detail, and is divided into two discrete half reactions that cycle between the PMP and PLP forms of the cofactor, overview
metabolism
physiological function
malfunction
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B-band LPS production is restored to Pseudomonas aeruginosa knockout mutants when the relevant Bordetella pertussis genes are supplied in trans
metabolism
additional information
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
purifed recombinant detagged WbpE in complex with the cofactor pyridoxal 5'-phosphate and product UDP-2-acetamido-3-amino-2,3-dideoxy-D-glucuronic acid as the external aldimine, mixing of 0.0015 ml of protein solution containing 10 mg/ml protein in 25 mM HEPES, pH 8.0, 100 mM NaCl, 0.5% glycerol, and 0.5 mM UDP-2-acetamido-3-amino-2,3-dideoxy-D-glucuronic acid, with 0.0015 ml of reservoir solution containing 0.1 M Bis-Tris, pH 5.5, 0.2 M ammonium sulfate and 25% PEG 3350 for the wild-type enzyme, and 0.1 M Bis-Tris, pH 5.5, 0.2 M ammonium acetate, 10 mM SrCl2 and 25% PEG 3350 for the sselenomethionine-labeled enzyme, X-ray diffraction structure determination and analysis at 1.9 A resolution, molecular replacement
purified enzyme in complex with dTDP-3-amino-3,6-dideoxy-D-galactose and dTDP-3-amino-3,6-dideoxy-D-glucose, X-ray diffraction structure determination and analysis at 1.3 A resolution, structure correction
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D156A
site-directed mutagenesis, the mutant shows only slightly affected activity and slight loss of binding affinity compared to the wild-type
H308A
site-directed mutagenesis, the mutant shows only slightly affected activity and slight loss of binding affinity compared to the wild-type
K185A
site-directed mutagenesis, the activity of the catalytic site mutant is completely abolished
N227A
site-directed mutagenesis, the mutant shows only slightly affected activity and slight loss of binding affinity compared to the wild-type
Q159A
site-directed mutagenesis, the mutant shows only slightly affected activity and slight loss of binding affinity compared to the wild-type
R229A
site-directed mutagenesis, the mutant shows only slightly affected activity and slight loss of binding affinity compared to the wild-type
S180A
site-directed mutagenesis, the mutant shows only slightly affected activity and slight loss of binding affinity compared to the wild-type
T60A
site-directed mutagenesis, the mutant shows only slightly affected activity and slight loss of binding affinity compared to the wild-type
Y309A
site-directed mutagenesis, the mutant shows only slightly affected activity and slight loss of binding affinity compared to the wild-type
additional information
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construction of a nonpolar knockout of each of wbpA, wbpB, wbpE, wbpD, and wbpI genes. Expression of Bordetella pertussis genes wbpO1629 and wbpO3150 complements a wbpA knockout of Pseudomonas aeruginosa. B-band LPS production is restored to Pseudomonas aeruginosa knockout mutants when the relevant Bordetella pertussis genes are supplied in trans
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20°C, purified recombinant His6-tagged WbpE, 3 weeks, stable
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precipitation occurs if the protein is left at ambient temperature or subjected to excessive freeze/thaw cycles
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PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
recombinant N-terminal His6-tagged WbpE from Escherichia coli BL21-CodonPlus(DE3)RIL by nickel affinity chromatography, the N-terminal His6-tag is removed by proteolysis with TEV protease over the course of three days while stirring in dialysis buffer, followed by gel filtration
recombinant His-tagged WpbE from Escherichia coli strain BL21-CodonPlus(DE3)RIL by nickel affinity chromatography
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recombinant His6-tagged WbpE from Escherichia coli strain Rosetta(DE3)
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CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
gene wbpE, expression of N-terminal His6-tagged WbpE in Escherichia coli BL21-CodonPlus(DE3)RIL
gene wbpE, expression of His6-tagged WbpE in Escherichia coli strain Rosetta(DE3)
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gene wbpE, expression of wild-type and mutants in Escherichia coli strain BL21(DE3)
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gene wpbE, overexpression of N-terminally T7- and C-terminally His6-tagged WpbE in Escherichia coli strain BL21-CodonPlus(DE3)RIL
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REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Westman, E.L.; Preston, A.; Field, R.A.; Lam, J.S.
Biosynthesis of a rare di-N-acetylated sugar in the lipopolysaccharides of both Pseudomonas aeruginosa and Bordetella pertussis occurs via an identical scheme despite different gene clusters
J. Bacteriol.
190
6060-6069
2008
Bordetella pertussis, Pseudomonas aeruginosa
Manually annotated by BRENDA team
Larkin, A.; Olivier, N.B.; Imperiali, B.
Structural analysis of WbpE from Pseudomonas aeruginosa PAO1: a nucleotide sugar aminotransferase involved in O-antigen assembly
Biochemistry
49
7227-7237
2010
Pseudomonas aeruginosa (Q9HZ76), Pseudomonas aeruginosa
Manually annotated by BRENDA team
Westman, E.L.; McNally, D.J.; Charchoglyan, A.; Brewer, D.; Field, R.A.; Lam, J.S.
Characterization of WbpB, WbpE, and WbpD and reconstitution of a pathway for the biosynthesis of UDP-2,3-diacetamido-2,3-dideoxy-D-mannuronic acid in Pseudomonas aeruginosa
J. Biol. Chem.
284
11854-11862
2009
Pseudomonas aeruginosa
Manually annotated by BRENDA team
Larkin, A.; Imperiali, B.
Biosynthesis of UDP-GlcNAc(3NAc)A by WbpB, WbpE, and WbpD: enzymes in the Wbp pathway responsible for O-antigen assembly in Pseudomonas aeruginosa PAO1
Biochemistry
48
5446-5455
2009
Pseudomonas aeruginosa
Manually annotated by BRENDA team
Dow, G.T.; Gilbert, M.; Thoden, J.B.; Holden, H.M.
Structural investigation on WlaRG from Campylobacter jejuni A sugar aminotransferase
Protein Sci.
26
586-599
2017
Pseudomonas aeruginosa (Q9HZ76), Pseudomonas aeruginosa 1C (Q9HZ76), Pseudomonas aeruginosa ATCC 15692 (Q9HZ76), Pseudomonas aeruginosa CIP 104116 (Q9HZ76), Pseudomonas aeruginosa DSM 22644 (Q9HZ76), Pseudomonas aeruginosa JCM 14847 (Q9HZ76), Pseudomonas aeruginosa LMG 12228 (Q9HZ76), Pseudomonas aeruginosa PRS 101 (Q9HZ76)
Manually annotated by BRENDA team