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Literature summary extracted from

  • Wenzel, C.Q.; Daniels, C.; Keates, R.A.; Brewer, D.; Lam, J.S.
    Evidence that WbpD is an N-acetyltransferase belonging to the hexapeptide acyltransferase superfamily and an important protein for O-antigen biosynthesis in Pseudomonas aeruginosa PAO1 (2005), Mol. Microbiol., 57, 1288-1303.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.3.1.5 gene wbpD, overexpression of the N-terminally His-tagged enzyme in strain CQW40 Pseudomonas aeruginosa
2.3.1.201 expressed as an N-terminally histidine-tagged fusion protein in Escherichia coli BL21(DE3) pLysS cells Pseudomonas aeruginosa

Protein Variants

EC Number Protein Variants Comment Organism
2.3.1.5 K136A site-directed mutagenesis, the mutant can only partially complement the wbpD knockout mutant strain, and shows also reduced stabilizing effects of acetyl-CoA on the mutant enzyme, while a K136R mutation shows no discernible effect on complementation of the wbpD mutant or the stabilizing effects of acetyl-CoA on the purified mutant protein Pseudomonas aeruginosa
2.3.1.5 K136R site-directed mutagenesis, partial complementation of the knockout mutant strain Pseudomonas aeruginosa
2.3.1.5 K58A site-directed mutagenesis, complete complementation of the knockout mutant strain to wild-type levels Pseudomonas aeruginosa
2.3.1.5 K58R site-directed mutagenesis, complete complementation of the knockout mutant strain to wild-type levels Pseudomonas aeruginosa
2.3.1.5 additional information construction of a chromosomal knockout mutant of wbpD is incapable of producing either long-chain B-band O antigen, with 2 or more repeating units, or semi-rough LPS, i.e. lipid A-core + one repeat, adding wbpD in trans restores LPS production to the wild-type level, overview Pseudomonas aeruginosa
2.3.1.5 Q60A site-directed mutagenesis, partial complementation of the knockout mutant strain Pseudomonas aeruginosa
2.3.1.5 Q60N site-directed mutagenesis, partial complementation of the knockout mutant strain Pseudomonas aeruginosa
2.3.1.201 K136A the mutation reduces the stabilizing effects of acetyl-CoA Pseudomonas aeruginosa
2.3.1.201 K136R the mutation showed no discernible effect the stabilizing effects of acetyl-CoA on the purified mutant protein Pseudomonas aeruginosa

General Stability

EC Number General Stability Organism
2.3.1.5 acetyl-CoA significantly enhances the stability of the purified recombinant protein and prevents precipitation over a course of 14 days Pseudomonas aeruginosa
2.3.1.201 incubation of the enzyme with acetyl-CoA significantly enhances the stability of the protein and prevents precipitation over a course of 14 days Pseudomonas aeruginosa

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.3.1.5 22736
-
 3 * 22736, sequence calculation, 3 * 23000, recombinant His-tagged enzyme, SDS-PAGE Pseudomonas aeruginosa
2.3.1.5 23000
-
 3 * 22736, sequence calculation, 3 * 23000, recombinant His-tagged enzyme, SDS-PAGE Pseudomonas aeruginosa
2.3.1.5 61000
-
 recombinant His-tagged enzyme, gel filtration Pseudomonas aeruginosa
2.3.1.201 22726
-
 3 * 22726, His-tagged enzyme, calculated from amino acid sequence Pseudomonas aeruginosa
2.3.1.201 22726
-
 3 * 22726, MALDI-TOF mass spectrometry Pseudomonas aeruginosa
2.3.1.201 23000
-
 3 * 23000, His-tagged enzyme, SDS-PAGE Pseudomonas aeruginosa
2.3.1.201 61000
-
 His-tagged enzyme, gel filtration Pseudomonas aeruginosa
2.3.1.201 68180
-
 MALDI-TOF mass spectrometry Pseudomonas aeruginosa

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.3.1.5 acetyl-CoA + UDP-2-acetamido-3-amino-2,3-dideoxy-D-glucuronic acid Pseudomonas aeruginosa WbpD is a putative 3-N-acetyltransferase that has been implicated in the biosynthesis of UDP-2,3-diacetamido-2,3-dideoxy-D-mannuronic acid, a precursor for the D-Man(2NAc3NAc)A residues in the B-band O antigen of Pseudomonas aeruginosa, the enzyme is involved in the catalysis of the fourth step by acting as an UDP-2-acetamido-3-amino-2,3-dideoxy-D-glucuronic acid 3-N-acetyltransferase CoA + UDP-2,3-diacetamido-2,3-dideoxy-D-glucuronic acid
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.3.1.5 Pseudomonas aeruginosa
-
 serotype O5, gene wbpD
-
2.3.1.201 Pseudomonas aeruginosa
-
 serotype O5
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.3.1.5 recombinant N-terminally His-tagged enzyme from strain CQW40 to over 95% purity by immobilized metal affinity chromatography Pseudomonas aeruginosa
2.3.1.201 Ni-NTA agarose column chromatography Pseudomonas aeruginosa

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.3.1.5 acetyl-CoA + UDP-2-acetamido-3-amino-2,3-dideoxy-D-glucuronic acid WbpD is a putative 3-N-acetyltransferase that has been implicated in the biosynthesis of UDP-2,3-diacetamido-2,3-dideoxy-D-mannuronic acid, a precursor for the D-Man(2NAc3NAc)A residues in the B-band O antigen of Pseudomonas aeruginosa, the enzyme is involved in the catalysis of the fourth step by acting as an UDP-2-acetamido-3-amino-2,3-dideoxy-D-glucuronic acid 3-N-acetyltransferase Pseudomonas aeruginosa CoA + UDP-2,3-diacetamido-2,3-dideoxy-D-glucuronic acid
-
 ?
2.3.1.5 acetyl-CoA + UDP-2-acetamido-3-amino-2,3-dideoxy-D-glucuronic acid WbpD utilizes a direct-transfer reaction mechanism Pseudomonas aeruginosa CoA + UDP-2,3-diacetamido-2,3-dideoxy-D-glucuronic acid
-
 ?
2.3.1.201 acetyl-CoA + UDP-2-acetamido-3-amino-2,3-dideoxy-alpha-D-glucuronate
-
 Pseudomonas aeruginosa CoA + UDP-2,3-diacetamido-2,3-dideoxy-alpha-D-glucuronate
-
 ?

Subunits

EC Number Subunits Comment Organism
2.3.1.5 More WbpD contains a left-handed beta-helical structure, secondary, tertiary, and quarternary structure determination by Far-UV circular dichroism spectroscopy analysis and crystal structure analysis, structure-based modeling, overview Pseudomonas aeruginosa
2.3.1.5 trimer 3 * 22736, sequence calculation, 3 * 23000, recombinant His-tagged enzyme, SDS-PAGE Pseudomonas aeruginosa
2.3.1.201 homotrimer 3 * 22726, His-tagged enzyme, calculated from amino acid sequence Pseudomonas aeruginosa
2.3.1.201 homotrimer 3 * 22726, MALDI-TOF mass spectrometry Pseudomonas aeruginosa
2.3.1.201 homotrimer 3 * 23000, His-tagged enzyme, SDS-PAGE Pseudomonas aeruginosa

Synonyms

EC Number Synonyms Comment Organism
2.3.1.5 More WbpD belongs to the hexapeptide acyltransferase superfamily of enzymes Pseudomonas aeruginosa
2.3.1.5 UDP-2-acetamido-3-amino-2,3-dideoxy-D-glucuronic acid 3-N-acetyltransferase
-
 Pseudomonas aeruginosa
2.3.1.5 UDP-D-Glc(2NAc3N)A 3-N-acetyltransferase
-
 Pseudomonas aeruginosa
2.3.1.5 WbpD
-
 Pseudomonas aeruginosa
2.3.1.201 UDP-2-acetamido-3-amino-2,3-dideoxy-D-glucuronic acid3-N-acetyltransferase
-
 Pseudomonas aeruginosa
2.3.1.201 WbpD
-
 Pseudomonas aeruginosa

Cofactor

EC Number Cofactor Comment Organism Structure
2.3.1.5 acetyl-CoA Glu60 and especially Lys136 play important roles in acetyl-CoA binding Pseudomonas aeruginosa

pI Value

EC Number Organism Comment pI Value Maximum pI Value
2.3.1.5 Pseudomonas aeruginosa recombinant enzyme, isoelectric focusing
-
 8
2.3.1.5 Pseudomonas aeruginosa sequence calculation
-
 8.22
2.3.1.201 Pseudomonas aeruginosa pI-value of above 8.0, isoelectric focusing
-
 8
2.3.1.201 Pseudomonas aeruginosa calculated from amino acid sequence
-
 8.2

General Information

EC Number General Information Comment Organism
2.3.1.201 physiological function the enzyme is required for B-band O-antigen biosynthesis in Pseudomonas aeruginosa Pseudomonas aeruginosa