Cloned (Comment) | Organism |
---|---|
gene pphC or yegK, DNA and amino acid sequence determination and analysis, sequence comparisons and phylogenetic analysis, recombinant expression of N-terminally His6-tagged wild-type and mutant enzymes in Escherichia coli strain C43 (DE3) | Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
D46N | site-directed mutagenesis, catalytically inactive mutant | Escherichia coli |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
5,5'-methylene disalicylic acid | 13% inhibition at 0.1 mM | Escherichia coli | |
Aurin tricarboxylic acid | 48% inhibition at 0.1 mM | Escherichia coli | |
EDTA | 62% inhibition at 2 mM | Escherichia coli | |
additional information | no inhibition by sanguinarine chloride at 0.05 mM. Poor inhibition by orthovanadate at 0.005-0.05 mM | Escherichia coli | |
okadaic acid | 12% inhibition at 0.001 mM | Escherichia coli | |
Sodium fluoride | 30% inhibition at 100 mM, 18% at 10 mM | Escherichia coli | |
Sodium phosphate | 87% inhibition at 5 mM, 96% at 10 mM | Escherichia coli |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | slightly activating | Escherichia coli | |
Mn2+ | highly activating, PphC (YegK) is a Mn2+-dependent PP2C-like phosphatase, best at 1-2 mM | Escherichia coli | |
additional information | no effect by Ca2+, Zn2+, or Ni2+ | Escherichia coli |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
[YegI kinase]-serine/threonine phosphate + H2O | Escherichia coli | dephosphorylation of autophosphorylated Escherichia coli YegI. Gene yegI encodes the partner kinase YegI of PphC | [YegI kinase]-serine/threonine + phosphate | - |
? | |
[YegI kinase]-serine/threonine phosphate + H2O | Escherichia coli MG1655 | dephosphorylation of autophosphorylated Escherichia coli YegI. Gene yegI encodes the partner kinase YegI of PphC | [YegI kinase]-serine/threonine + phosphate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | P76395 | - |
- |
Escherichia coli MG1655 | P76395 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant N-terminally His6-tagged wild-type and mutant enzymes from Escherichia coli strain C43 (DE3) by nickel affinity chromatography | Escherichia coli |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | PphC displays low preference for pSer/pThr/pTyr peptides, the minimal activity against phosphopeptides is in contrast to the ability of PphC to dephosphorylate the pSer-containing protein substrate beta-casein. PphC may require additional residues for substrate recognition and/or binding that are not present in the phosphopeptides | Escherichia coli | ? | - |
? | |
additional information | PphC displays low preference for pSer/pThr/pTyr peptides, the minimal activity against phosphopeptides is in contrast to the ability of PphC to dephosphorylate the pSer-containing protein substrate beta-casein. PphC may require additional residues for substrate recognition and/or binding that are not present in the phosphopeptides | Escherichia coli MG1655 | ? | - |
? | |
[YegI kinase]-serine/threonine phosphate + H2O | dephosphorylation of autophosphorylated Escherichia coli YegI. Gene yegI encodes the partner kinase YegI of PphC | Escherichia coli | [YegI kinase]-serine/threonine + phosphate | - |
? | |
[YegI kinase]-serine/threonine phosphate + H2O | dephosphorylation of autophosphorylated Escherichia coli YegI. Gene yegI encodes the partner kinase YegI of PphC | Escherichia coli MG1655 | [YegI kinase]-serine/threonine + phosphate | - |
? |
Subunits | Comment | Organism |
---|---|---|
? | x * 25000, recombinant His6-tagged enzyme, SDS-PAGE | Escherichia coli |
Synonyms | Comment | Organism |
---|---|---|
phosphatase 2C-like Ser/Thr phosphatase | - |
Escherichia coli |
PP2C-like phosphatase | - |
Escherichia coli |
PphC | - |
Escherichia coli |
YegK | - |
Escherichia coli |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | 37 | assay at | Escherichia coli |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | 8 | assay at | Escherichia coli |
General Information | Comment | Organism |
---|---|---|
evolution | enzyme PphC belongs to the eukaryote-like Ser/Thr phosphatases (eSTPs) in Escherichia coli, that have extensive sequence and structural homology to eukaryotic Ser/Thr protein phosphatase 2C (PP2C) phosphatases. But YegK is an atypical PP2C-like phosphatase. Unlike other bacterial PP2C homologues, YegK contains only six of the eight absolutely conserved residues that are involved in metal binding, coordination, and catalysis, instead of eleven. In particular, the amino acid sequence alignment clearly shows that YegK lacks the conserved glycine residue in motif VI and the aspartic acid residue in motif VIII | Escherichia coli |
physiological function | the Escherichia coli eSTP acts to dephosphorylate another Ser/Thr kinase that is encoded in the same operon. Regulatory reversible protein phosphorylation is a conserved mechanism of signaling in all biological systems | Escherichia coli |