Cloned (Comment) | Organism |
---|---|
gene wbpD, overexpression of the N-terminally His-tagged enzyme in strain CQW40 | Pseudomonas aeruginosa |
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 | 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 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 | 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 | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas aeruginosa | - |
serotype O5, gene wbpD | - |
Purification (Comment) | Organism |
---|---|
recombinant N-terminally His-tagged enzyme from strain CQW40 to over 95% purity by immobilized metal affinity chromatography | Pseudomonas aeruginosa |
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 | 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 | 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 | Comment | Organism | Structure |
---|---|---|---|
acetyl-CoA | Glu60 and especially Lys136 play important roles in acetyl-CoA binding | Pseudomonas aeruginosa |
Organism | Comment | pI Value Maximum | pI Value |
---|---|---|---|
Pseudomonas aeruginosa | recombinant enzyme, isoelectric focusing | - |
8 |
Pseudomonas aeruginosa | sequence calculation | - |
8.22 |