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Literature summary for 2.7.8.24 extracted from
- Enzymatic properties and substrate specificity of a bacterial phosphatidylcholine synthase (2014), FEBS J., 281, 3523-3541.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| recombinant expression of C-terminally His6-tagged enzyme in Escherichia coli, recombinant expression of His6-tagged enzyme mutants in Escherichia coli strain C41 (DE3) | Agrobacterium tumefaciens |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| D100A | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Agrobacterium tumefaciens |
| D104A | site-directed mutagenesis, inactive mutant | Agrobacterium tumefaciens |
| D78A | site-directed mutagenesis, almost inactive mutant | Agrobacterium tumefaciens |
| G79A | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Agrobacterium tumefaciens |
| G96A | site-directed mutagenesis, the mutant shows activity similar to the wild-type enzyme | Agrobacterium tumefaciens |
| R83A | site-directed mutagenesis, the mutant shows activity similar to the wild-type enzyme | Agrobacterium tumefaciens |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| carnitine | inhibits the enzyme's activity in vitro | Agrobacterium tumefaciens |  |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| additional information | - |
additional information | choline-binding by the enzyme is cooperative | Agrobacterium tumefaciens |
Localization
| Localization | Comment | Organism | GeneOntology No. | Textmining |
|---|---|---|---|---|
| membrane | the enzyme is an integral membrane protein, that consists of eight transmembrane segments with its N- and C-termini located in the cytoplasm, membrane topology of the enzyme, reaction mechanism, overview | Agrobacterium tumefaciens | 16020 | - |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Mn2+ | required for activity | Agrobacterium tumefaciens | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 400000 | - |
His6-tagged recombinant enzyme, gel filtration | Agrobacterium tumefaciens |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| CDP-diacylglycerol + choline | Agrobacterium tumefaciens | - |
CMP + phosphatidylcholine | - |
r | |
| CDP-diacylglycerol + choline | Agrobacterium tumefaciens C58 / ATCC 33970 | - |
CMP + phosphatidylcholine | - |
r | |
| additional information | Agrobacterium tumefaciens | no activity with gamma-butyrobetaine, betaine, and carnitine instead of choline | ? | - |
? | |
| additional information | Agrobacterium tumefaciens C58 / ATCC 33970 | no activity with gamma-butyrobetaine, betaine, and carnitine instead of choline | ? | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Agrobacterium tumefaciens | A9CIM3 | - |
- |
| Agrobacterium tumefaciens C58 / ATCC 33970 | A9CIM3 | - |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant C-terminally His6-tagged enzyme from Escherichia coli by nickel affinity chromatography, recombinant His6-tagged enzyme mutants from Escherichia coli strain C41 (DE3) | Agrobacterium tumefaciens |
Reaction
| Reaction | Comment | Organism | Reaction ID |
|---|---|---|---|
| CDP-diacylglycerol + choline = CMP + phosphatidylcholine | the enzyme reaction follows a reversible two-step reaction: in a first choline- and manganese-independent reaction, CDP-diacylglycerol is hydrolyzed releasing a CMP molecule. The resulting phosphatidyl intermediate reacts with choline in a second manganese-dependent step to form phosphatidylcholine | Agrobacterium tumefaciens |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| CDP-diacylglycerol + choline | - |
Agrobacterium tumefaciens | CMP + phosphatidylcholine | - |
r | |
| CDP-diacylglycerol + choline | phosphatidylcholine formation by enzyme Pcs is reversible and proceeds via two successive reactions | Agrobacterium tumefaciens | CMP + phosphatidylcholine | - |
r | |
| CDP-diacylglycerol + choline | - |
Agrobacterium tumefaciens C58 / ATCC 33970 | CMP + phosphatidylcholine | - |
r | |
| CDP-diacylglycerol + choline | phosphatidylcholine formation by enzyme Pcs is reversible and proceeds via two successive reactions | Agrobacterium tumefaciens C58 / ATCC 33970 | CMP + phosphatidylcholine | - |
r | |
| additional information | no activity with gamma-butyrobetaine, betaine, and carnitine instead of choline | Agrobacterium tumefaciens | ? | - |
? | |
| additional information | the enzyme exhibits broad substrate specificity towards choline derivatives. The presence of CDP-diacylglycerol and manganese is a prerequisite for cooperative binding of choline. No activity with gamma-butyrobetaine, betaine, and carnitine instead of choline | Agrobacterium tumefaciens | ? | - |
? | |
| additional information | no activity with gamma-butyrobetaine, betaine, and carnitine instead of choline | Agrobacterium tumefaciens C58 / ATCC 33970 | ? | - |
? | |
| additional information | the enzyme exhibits broad substrate specificity towards choline derivatives. The presence of CDP-diacylglycerol and manganese is a prerequisite for cooperative binding of choline. No activity with gamma-butyrobetaine, betaine, and carnitine instead of choline | Agrobacterium tumefaciens C58 / ATCC 33970 | ? | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| oligomer | x * 23000-30000, SDS-PAGE and sequence calculation, the enzyme forms stable oligomers | Agrobacterium tumefaciens |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| PC synthase | - |
Agrobacterium tumefaciens |
| PCS | - |
Agrobacterium tumefaciens |
General Information
| General Information | Comment | Organism |
|---|---|---|
| additional information | cytoplasmic loop between the second and third membrane helix contains the majority of the conserved amino acids of a CDP-alcohol phosphotransferase motif (DGX2ARX12GX3DX3D), crucial role of this motif in choline binding and enzyme activity | Agrobacterium tumefaciens |
| physiological function | phosphatidylcholine, produced by the enzyme, is essential for the expression of virulence-related genes that is controlled by the two-component system VirA/G | Agrobacterium tumefaciens |
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