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Literature summary for 3.1.4.3 extracted from
- The Legionella pneumophila Dot/Icm-secreted effector PlcC/CegC1 together with PlcA and PlcB promotes virulence and belongs to a novel zinc metallophospholipase C family present in bacteria and fungi (2013), J. Biol. Chem., 288, 11080-11092.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| gene lpg0012 or plcC, DNA and amino acid sequence determination and analysis, recombinant expression of N-terminally His6-tagged enzyme in Escherichia coli strain BL21, subcloning in Escherichia coli strain DH5alpha. Expression of Strep-tagged enzyme mutants H179N and S336A does not yield any detectable protein | Legionella pneumophila |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| D251V | site-directed mutagenesis, the mutant enzyme shows reduced catalytic activity compared to the wild-type enzyme | Legionella pneumophila |
| D314V | site-directed mutagenesis, the mutant enzyme shows reduced catalytic activity compared to the wild-type enzyme | Legionella pneumophila |
| D63V | site-directed mutagenesis, the mutant enzyme shows reduced catalytic activity compared to the wild-type enzyme | Legionella pneumophila |
| E286A | site-directed mutagenesis, the mutant enzyme shows reduced catalytic activity compared to the wild-type enzyme | Legionella pneumophila |
| F167A | site-directed mutagenesis, the mutant enzyme shows reduced catalytic activity compared to the wild-type enzyme | Legionella pneumophila |
| F244A | site-directed mutagenesis, the mutant enzyme shows reduced catalytic activity compared to the wild-type enzyme | Legionella pneumophila |
| F253A | site-directed mutagenesis, the mutant enzyme shows reduced catalytic activity compared to the wild-type enzyme | Legionella pneumophila |
| H166N | site-directed mutagenesis, the mutant enzyme shows reduced catalytic activity compared to the wild-type enzyme | Legionella pneumophila |
| H179N | site-directed mutagenesis, the mutant enzyme shows reduced catalytic activity compared to the wild-type enzyme | Legionella pneumophila |
| H247N | site-directed mutagenesis, the mutant enzyme shows reduced catalytic activity compared to the wild-type enzyme | Legionella pneumophila |
| H257N | site-directed mutagenesis, the mutant enzyme shows reduced catalytic activity compared to the wild-type enzyme | Legionella pneumophila |
| H284N | site-directed mutagenesis, the mutant enzyme shows reduced catalytic activity compared to the wild-type enzyme | Legionella pneumophila |
| H409N | site-directed mutagenesis, the mutant enzyme shows similar catalytic activity as the wild-type enzyme | Legionella pneumophila |
| additional information | construction of enzyme knock-out mutants, of PlcC alone, or double PlcC/PlcA or PlcC/PlcB and triple PlcC/PlcA/PlcB mutants. Complementation of plcC knock-out mutants with wild-type plcC in trans restores the cell-associated enzyme activity defect | Legionella pneumophila |
| R265Q | site-directed mutagenesis, the mutant enzyme shows reduced catalytic activity compared to the wild-type enzyme | Legionella pneumophila |
| R326Q | site-directed mutagenesis, the mutant enzyme shows reduced catalytic activity compared to the wild-type enzyme | Legionella pneumophila |
| R365Q | site-directed mutagenesis, the mutant enzyme shows reduced catalytic activity compared to the wild-type enzyme | Legionella pneumophila |
| S336A | site-directed mutagenesis, the mutant enzyme shows reduced catalytic activity compared to the wild-type enzyme | Legionella pneumophila |
| Y156A | site-directed mutagenesis, the mutant enzyme shows reduced catalytic activity compared to the wild-type enzyme | Legionella pneumophila |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| EDTA | - |
Legionella pneumophila |  |
Localization
| Localization | Comment | Organism | GeneOntology No. | Textmining |
|---|---|---|---|---|
| extracellular | IVB Dot/Icm secretion type. The secreted enzyme activity depends upon an intact type II secretion system | Legionella pneumophila | - |
- |
| additional information | the enzyme sequence contains no signal peptide | Legionella pneumophila | - |
- |
| type IV secretion system complex | - |
Legionella pneumophila | 43684 | - |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Zn2+ | activates | Legionella pneumophila | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 60090 | - |
x * 60090, sequence calculation | Legionella pneumophila |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| phosphatidylcholine + H2O | Legionella pneumophila | the enzyme shows cell-associated phosphatidylcholine-specific phospholipase C activity in vivo | 1,2-sn-diacylglycerol + phosphocholine | - |
? | |
| phosphatidylcholine + H2O | Legionella pneumophila JR32 | the enzyme shows cell-associated phosphatidylcholine-specific phospholipase C activity in vivo | 1,2-sn-diacylglycerol + phosphocholine | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Legionella pneumophila | Q5ZZJ8 | gene lpg0012 or plcC | - |
| Legionella pneumophila JR32 | Q5ZZJ8 | gene lpg0012 or plcC | - |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| additional information | the recombinant enzyme hydrolyzes a broad phospholipid spectrum, including phosphatidylcholine, phosphatidylglycerol, and phosphatidylinositol, broad substrate preference | Legionella pneumophila | ? | - |
? | |
| additional information | the recombinant enzyme hydrolyzes a broad phospholipid spectrum, including phosphatidylcholine, phosphatidylglycerol, and phosphatidylinositol, broad substrate preference | Legionella pneumophila JR32 | ? | - |
? | |
| phosphatidylcholine + H2O | - |
Legionella pneumophila | 1,2-sn-diacylglycerol + phosphocholine | - |
? | |
| phosphatidylcholine + H2O | the enzyme shows cell-associated phosphatidylcholine-specific phospholipase C activity in vivo | Legionella pneumophila | 1,2-sn-diacylglycerol + phosphocholine | - |
? | |
| phosphatidylcholine + H2O | - |
Legionella pneumophila JR32 | 1,2-sn-diacylglycerol + phosphocholine | - |
? | |
| phosphatidylcholine + H2O | the enzyme shows cell-associated phosphatidylcholine-specific phospholipase C activity in vivo | Legionella pneumophila JR32 | 1,2-sn-diacylglycerol + phosphocholine | - |
? | |
| phosphatidylglycerol + H2O | - |
Legionella pneumophila | 1,2-sn-diacylglycerol + sn-glycerol-3-phosphate | - |
? | |
| phosphatidylglycerol + H2O | - |
Legionella pneumophila JR32 | 1,2-sn-diacylglycerol + sn-glycerol-3-phosphate | - |
? | |
| phosphatidylinositol + H2O | - |
Legionella pneumophila | 1,2-sn-diacylglycerol + phosphoinositol | - |
? | |
| phosphatidylinositol + H2O | - |
Legionella pneumophila JR32 | 1,2-sn-diacylglycerol + phosphoinositol | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| ? | x * 60090, sequence calculation | Legionella pneumophila |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| CegC1 | - |
Legionella pneumophila |
| Dot/Icm-injected effector | - |
Legionella pneumophila |
| Lpg0012 | - |
Legionella pneumophila |
| PC-PLC | - |
Legionella pneumophila |
| phosphatidylcholine-specific phospholipase C | - |
Legionella pneumophila |
| plcC | - |
Legionella pneumophila |
| PlcC/CegC1 | - |
Legionella pneumophila |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 37 | - |
assay at | Legionella pneumophila |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7.5 | - |
assay at | Legionella pneumophila |
pI Value
| Organism | Comment | pI Value Maximum | pI Value |
|---|---|---|---|
| Legionella pneumophila | sequence calculation | - |
6.55 |
Expression
| Organism | Comment | Expression |
|---|---|---|
| Legionella pneumophila | the enzyme is upregulated during infection of Acanthamoeba castellanii amoebae and macrophages | up |
General Information
| General Information | Comment | Organism |
|---|---|---|
| evolution | the enzyme belongs to a distinct zinc metallophospholipase C family present in bacteria and fungi | Legionella pneumophila |
| malfunction | enzyme knock-out mutants of PlcC show abolished enzyme activity. Complementation of plcC knock-out mutants with wild-type plcC in trans restores the cell-associated enzyme activity defect | Legionella pneumophila |
| additional information | the fifteen conserved amino acids Asp63, Tyr156, His166, Phe167, Phe244, His247, Asp251, Phe253, Arg265, His257, His284, Glu286, Asp314, Arg326, and Arg385 are essential for enzyme activity | Legionella pneumophila |
| physiological function | the enzyme, phosphatidylcholine-specific phospholipase C or effector PlcC/CegC1, together with Zn2+-dependent enzymes PlcA and PlcB, exhibiting phosphatidylglycerol hydrolysis activity, promote virulence, e.g. in Acanthamoeba castellanii amoebae and human U937 cells or in Galleria mellonella larvae, but are not essential. The enzyme is CpxR-co-regulated | Legionella pneumophila |
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