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Literature summary for 2.3.1.5 extracted from
- 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.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| gene wbpD, overexpression of the N-terminally His-tagged enzyme in strain CQW40 | Pseudomonas aeruginosa |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| 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 |
| K136R | site-directed mutagenesis, partial complementation of the knockout mutant strain | Pseudomonas aeruginosa |
| K58A | site-directed mutagenesis, complete complementation of the knockout mutant strain to wild-type levels | Pseudomonas aeruginosa |
| K58R | site-directed mutagenesis, complete complementation of the knockout mutant strain to wild-type levels | Pseudomonas aeruginosa |
| 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 |
| Q60A | site-directed mutagenesis, partial complementation of the knockout mutant strain | Pseudomonas aeruginosa |
| Q60N | site-directed mutagenesis, partial complementation of the knockout mutant strain | Pseudomonas aeruginosa |
General Stability
| General Stability | Organism |
|---|---|
| acetyl-CoA significantly enhances the stability of the purified recombinant protein and prevents precipitation over a course of 14 days | Pseudomonas aeruginosa |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 22736 | - |
3 * 22736, sequence calculation, 3 * 23000, recombinant His-tagged enzyme, SDS-PAGE | Pseudomonas aeruginosa |
| 23000 | - |
3 * 22736, sequence calculation, 3 * 23000, recombinant His-tagged enzyme, SDS-PAGE | Pseudomonas aeruginosa |
| 61000 | - |
recombinant His-tagged enzyme, gel filtration | Pseudomonas aeruginosa |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 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
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Pseudomonas aeruginosa | - |
serotype O5, gene wbpD | - |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant N-terminally His-tagged enzyme from strain CQW40 to over 95% purity by immobilized metal affinity chromatography | Pseudomonas aeruginosa |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 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 | - |
? | |
| 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 | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| 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 |
| trimer | 3 * 22736, sequence calculation, 3 * 23000, recombinant His-tagged enzyme, SDS-PAGE | Pseudomonas aeruginosa |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| More | WbpD belongs to the hexapeptide acyltransferase superfamily of enzymes | Pseudomonas aeruginosa |
| UDP-2-acetamido-3-amino-2,3-dideoxy-D-glucuronic acid 3-N-acetyltransferase | - |
Pseudomonas aeruginosa |
| UDP-D-Glc(2NAc3N)A 3-N-acetyltransferase | - |
Pseudomonas aeruginosa |
| WbpD | - |
Pseudomonas aeruginosa |
Cofactor
| Cofactor | Comment | Organism | Structure |
|---|---|---|---|
| acetyl-CoA | Glu60 and especially Lys136 play important roles in acetyl-CoA binding | Pseudomonas aeruginosa | |
pI Value
| Organism | Comment | pI Value Maximum | pI Value |
|---|---|---|---|
| Pseudomonas aeruginosa | recombinant enzyme, isoelectric focusing | - |
8 |
| Pseudomonas aeruginosa | sequence calculation | - |
8.22 |
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