Cloned (Comment) | Organism |
---|---|
gene acs, recombinant expression of His-tagged wild-type and mutant enzymes in Salmonella enterica strain G2466 | Salmonella enterica |
Crystallization (Comment) | Organism |
---|---|
purified recombinant enzyme SeAcs in complex with cAMP and CoA, X-ray diffraction structure determination and analysis at 1.65 A resolution | Salmonella enterica |
Protein Variants | Comment | Organism |
---|---|---|
D411A | site-directed mutagenesis, the mutant SeAcs variant shows a nearly 10fold increased dissociation constant compared to the wild-type enzyme | Salmonella enterica |
D500A | site-directed mutagenesis, the mutant SeAcs variant is defective in cAMP binding | Salmonella enterica |
additional information | cAMP has no effect on acetylation promotion of the mutant SeAcs enzymes | Salmonella enterica |
N521A | site-directed mutagenesis, the mutant SeAcs variant shows a higher dissociation constant compared to the wild-type enzyme | Salmonella enterica |
Q415A | site-directed mutagenesis, the mutant SeAcs variant is defective in cAMP binding | Salmonella enterica |
R515A | site-directed mutagenesis, the mutant SeAcs variant shows a nearly 10fold increased dissociation constant compared to the wild-type enzyme | Salmonella enterica |
T416A | site-directed mutagenesis, the mutant SeAcs variant shows a higher dissociation constant compared to the wild-type enzyme | Salmonella enterica |
W413A | site-directed mutagenesis, the mutant SeAcs variant is defective in cAMP binding | Salmonella enterica |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
cAMP | cyclic AMP inhibits the activity and promotes the acetylation of acetyl-CoA synthetase through competitive binding to the highly conserved ATP/AMP binding pocket and restrains SeAcs in an open conformation. cAMP directly binds to the enzyme and inhibits its activity in a substrate-competitive manner. cAMP binding increases SeAcs acetylation by simultaneously promoting Pat-dependent acetylation and inhibiting CobB-dependent deacetylation, resulting in enhanced SeAcs inhibition | Salmonella enterica |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | reuqired | Salmonella enterica |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + acetate + CoA | Salmonella enterica | - |
AMP + diphosphate + acetyl-CoA | - |
? | |
ATP + acetate + CoA | Salmonella enterica G2466 | - |
AMP + diphosphate + acetyl-CoA | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Salmonella enterica | - |
- |
- |
Salmonella enterica G2466 | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged wild-type and mutant enzymes from Salmonella enterica strain G2466 by nickel affinity chromatography | Salmonella enterica |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + acetate + CoA | - |
Salmonella enterica | AMP + diphosphate + acetyl-CoA | - |
? | |
ATP + acetate + CoA | - |
Salmonella enterica G2466 | AMP + diphosphate + acetyl-CoA | - |
? | |
additional information | activity determination in a coupled assay with myokinase, pyruvate kinase, and lactate dehydrogenase: SeAcs first converts acetate, CoA, and ATP to acetyl-CoA and AMP. Then, myokinase converts AMP to ADP. Pyruvate kinase converts ADP and phosphoenolpyruvate to pyruvate and ATP. Finally, lactate dehydrogenase reduces pyruvate and oxidizes NADH to NAD+ | Salmonella enterica | ? | - |
? | |
additional information | activity determination in a coupled assay with myokinase, pyruvate kinase, and lactate dehydrogenase: SeAcs first converts acetate, CoA, and ATP to acetyl-CoA and AMP. Then, myokinase converts AMP to ADP. Pyruvate kinase converts ADP and phosphoenolpyruvate to pyruvate and ATP. Finally, lactate dehydrogenase reduces pyruvate and oxidizes NADH to NAD+ | Salmonella enterica G2466 | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
Acetyl-CoA synthetase | - |
Salmonella enterica |
acetyl-coenzyme A synthetase | - |
Salmonella enterica |
ACS | - |
Salmonella enterica |
SeAcs | - |
Salmonella enterica |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Salmonella enterica |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Salmonella enterica |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | - |
Salmonella enterica |
General Information | Comment | Organism |
---|---|---|
malfunction | cyclic AMP inhibits the activity and promotes the acetylation of acetyl-CoA synthetase through competitive binding to the ATP/AMP pocket. cAMP directly binds to the enzyme and inhibits its activity in a substrate-competitive manner. cAMP binding increases SeAcs acetylation by simultaneously promoting Pat-dependent acetylation and inhibiting CobB-dependent deacetylation, resulting in enhanced SeAcs inhibition | Salmonella enterica |
physiological function | the high-affinity biosynthetic pathway for converting acetate to acetyl-coenzyme A (acetyl-CoA) is catalyzed by the central metabolic enzyme acetyl-coenzyme A synthetase (Acs), which is finely regulated both at the transcriptional level via cyclic AMP (cAMP)-driven trans-activation and at the post-translational level via acetylation inhibition. The cAMP contact residues are well conserved from prokaryotes to eukaryotes, suggesting a general regulatory mechanism of cAMP on Acs. cAMP generally inhibits other acyl- or aryl-CoA synthetases | Salmonella enterica |