Cloned (Comment) | Organism |
---|---|
gene PA4534, recombinant expression of N-terminally His-tagged enzyme in Escherichia coli strain BL21(DE3) | Pseudomonas aeruginosa |
Crystallization (Comment) | Organism |
---|---|
purified recombinant detagged enzyme in apoform and bound to acetyl-CoA, hanging drop vapor diffusion method, mixing of 8.5 mg/ml protein solution with reservoir solution contains 16% PEG 1000, 0.2 M calcium acetate, and 0.1 M Tris-HCl, pH 7.0, crystals of acetyl-CoA bound enzyme are obtained by mixing 0.49 mM protein and 0.7 mM acetyl-CoA and equilibrating with a well solution composed of 15% PEG 3350, 0.2 M ammonium citrate dibasic, and 0.1 M Tris-HCl, pH 7.5, 20°C, X-ray diffraction structure determination and analysis at 2.21 A and 1.62 A resolution, respectively, molecular replacement using the putative acetyltransferase YpeA structure (PDB ID 2PDO) as a search model, model building | Pseudomonas aeruginosa |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas aeruginosa | Q9HVP3 | - |
- |
Pseudomonas aeruginosa 1C | Q9HVP3 | - |
- |
Pseudomonas aeruginosa ATCC 15692 | Q9HVP3 | - |
- |
Pseudomonas aeruginosa CIP 104116 | Q9HVP3 | - |
- |
Pseudomonas aeruginosa DSM 22644 | Q9HVP3 | - |
- |
Pseudomonas aeruginosa JCM 14847 | Q9HVP3 | - |
- |
Pseudomonas aeruginosa LMG 12228 | Q9HVP3 | - |
- |
Pseudomonas aeruginosa PRS 101 | Q9HVP3 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant N-terminally His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, tag cleavage through TEV protease, gel filtration, and ultrafiltration | Pseudomonas aeruginosa |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
acetyl-CoA + [protein]-L-lysine | - |
Pseudomonas aeruginosa | CoA + [protein]-N6-acetyl-L-lysine | - |
? | |
acetyl-CoA + [protein]-L-lysine | - |
Pseudomonas aeruginosa ATCC 15692 | CoA + [protein]-N6-acetyl-L-lysine | - |
? | |
acetyl-CoA + [protein]-L-lysine | - |
Pseudomonas aeruginosa 1C | CoA + [protein]-N6-acetyl-L-lysine | - |
? | |
acetyl-CoA + [protein]-L-lysine | - |
Pseudomonas aeruginosa PRS 101 | CoA + [protein]-N6-acetyl-L-lysine | - |
? | |
acetyl-CoA + [protein]-L-lysine | - |
Pseudomonas aeruginosa DSM 22644 | CoA + [protein]-N6-acetyl-L-lysine | - |
? | |
acetyl-CoA + [protein]-L-lysine | - |
Pseudomonas aeruginosa CIP 104116 | CoA + [protein]-N6-acetyl-L-lysine | - |
? | |
acetyl-CoA + [protein]-L-lysine | - |
Pseudomonas aeruginosa LMG 12228 | CoA + [protein]-N6-acetyl-L-lysine | - |
? | |
acetyl-CoA + [protein]-L-lysine | - |
Pseudomonas aeruginosa JCM 14847 | CoA + [protein]-N6-acetyl-L-lysine | - |
? |
Subunits | Comment | Organism |
---|---|---|
homodimer | the dimer interface includes a domain swapping where strand beta7 of subunit A is located between strands beta5 and beta6 of subunit B, forming an antiparallel beta-sheet | Pseudomonas aeruginosa |
Synonyms | Comment | Organism |
---|---|---|
N-acetyltransferase | - |
Pseudomonas aeruginosa |
PA4534 | - |
Pseudomonas aeruginosa |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
acetyl-CoA | - |
Pseudomonas aeruginosa |
General Information | Comment | Organism |
---|---|---|
evolution | the enzyme belongs to the GCN5-related N-acetyltransferase (GNAT) superfamily. The GNAT superfamily of proteins is involved in many physiologically important reactions in eukaryotes and prokaryotes. PA4534 shares common characteristic structures with other GNAT family N-acetyltransferases and contains a potential substrate binding tunnel close to the bound acetyl-CoA. It also shows the characteristic features of GNAT proteins including the beta-bulge in b4 and the P-loop between beta4 and alpha3. The P-loop of PA4534 locates between beta4 and alpha3 that connects the diphosphate group of acetyl-CoA with the main chain nitrogen atoms from residues including G81, G83, A85, and R86 | Pseudomonas aeruginosa |
additional information | the enzyme contains an acetyl-CoA binding site located at the V-shaped cleft between beta4-beta5 strands created by beta-bulge on beta4. Another possible substrate binding site is identified close to the acetyl group of bound acetyl-CoA molecule. Acetyl-CoA adopts a C-shaped conformation where the adenosine diphosphate moiety is partly exposed to solvent and the acetyl group that is transferred to a substrate in a N-acetyltransferase reaction is deeply buried in the protein pointing towards a tunnel, potential substrate binding tunnel close to acetyl-CoA, overview. Structure comparisons | Pseudomonas aeruginosa |
physiological function | GCN5-related N-acetyltransferases (GNATs) are a large and diverse group of enzymes which catalyze the transfer of an acetyl group from acetyl coenzyme A (Ac-CoA) to the amine group of a substrate. Substrates include protein N-terminus, lysine of histone tails, and other small molecules such as aminoglycoside, serotonin, and glucose-6-phosphate | Pseudomonas aeruginosa |