Cloned (Comment) | Organism |
---|---|
gene prpC, sequence comparisons, recombinant expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3). The presence of the His-tag does not disturb the overall protein secondary structure, even when metal is added | Mycoplasmopsis synoviae |
Protein Variants | Comment | Organism |
---|---|---|
D122A | site-directed mutagenesis, catalytically inactive mutant | Mycoplasmopsis synoviae |
R164A | site-directed mutagenesis, the mutant's catalytic efficiency is 2.3fold lower compared to wild-type enzyme | Mycoplasmopsis synoviae |
General Stability | Organism |
---|---|
metal binding contributes to the protein overall structural stability | Mycoplasmopsis synoviae |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
Ca2+ | - |
Mycoplasmopsis synoviae | |
Cu2+ | - |
Mycoplasmopsis synoviae | |
Zn2+ | strong inhibition, the inhibition of PrpC activity by Zn2+ ions may be attributed to the destabilization of the native fold of the protein | Mycoplasmopsis synoviae |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | Michaelis-Menten kinetics, steady-state kinetics, thermodynamics and isothermal titration calorimetry | Mycoplasmopsis synoviae | |
0.023 | - |
[alpha-casein]-serine/threonine phosphate | pH 9.0, 30°C, recombinant wild-type enzyme | Mycoplasmopsis synoviae | |
0.308 | - |
RRA(pT)VA | pH 9.0, 30°C, recombinant wild-type enzyme | Mycoplasmopsis synoviae |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | activates, can partially substitute for Mn2+ | Mycoplasmopsis synoviae | |
Mn2+ | dependent on, required for enzyme activity | Mycoplasmopsis synoviae | |
additional information | the conserved residues involved in PP2C third metal ion-binding site are also involved in PrpC catalysis, and the metal binding also contributes to the protein overall structural stability. Addition of 0.2 mM MnCl2 and 0.01 mM ZnSO4 to the enzymes does not induce large structural changes in PrpC, but at 0.06 mM ZnSO4 a strong effect in the secondary structure of PrpC is observed | Mycoplasmopsis synoviae |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
29000 | 30087 | recombinant His6-tagged wild-type enzyme, mass spectrometry and gel filtration | Mycoplasmopsis synoviae |
29000 | 29997 | recombinant His6-tagged mutant R164A, mass spectrometry and gel filtration | Mycoplasmopsis synoviae |
29000 | 30045 | recombinant His6-tagged mutant D122A, mass spectrometry and gel filtration | Mycoplasmopsis synoviae |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Mycoplasmopsis synoviae | Q4A6S8 | - |
- |
Mycoplasmopsis synoviae 53 | Q4A6S8 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, cleavage of the His-tag with thrombin, and gel filtration | Mycoplasmopsis synoviae |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
KR(pT)IRR + H2O | - |
Mycoplasmopsis synoviae | KRTIRR + phosphate | - |
? | |
KR(pT)IRR + H2O | - |
Mycoplasmopsis synoviae 53 | KRTIRR + phosphate | - |
? | |
additional information | enzyme PrpC has phosphatase activity in vitro, being able to dephosphorylate the artificial substrate 4-nitrophenyl phosphate, and phospho-Ser/Thr residues in peptides, as well as phospho-alpha-casein | Mycoplasmopsis synoviae | ? | - |
? | |
additional information | enzyme PrpC has phosphatase activity in vitro, being able to dephosphorylate the artificial substrate 4-nitrophenyl phosphate, and phospho-Ser/Thr residues in peptides, as well as phospho-alpha-casein | Mycoplasmopsis synoviae 53 | ? | - |
? | |
RRA(pS)VA + H2O | - |
Mycoplasmopsis synoviae | RRASVA + phosphate | - |
? | |
RRA(pS)VA + H2O | - |
Mycoplasmopsis synoviae 53 | RRASVA + phosphate | - |
? | |
RRA(pT)VA + H2O | - |
Mycoplasmopsis synoviae | RRATVA + phosphate | - |
? | |
RRA(pT)VA + H2O | - |
Mycoplasmopsis synoviae 53 | RRATVA + phosphate | - |
? | |
RRLIEDAE(pY)AARG + H2O | - |
Mycoplasmopsis synoviae | RRLIEDAEYAARG + phosphate | - |
? | |
RRLIEDAE(pY)AARG + H2O | - |
Mycoplasmopsis synoviae 53 | RRLIEDAEYAARG + phosphate | - |
? | |
[alpha-casein]-serine/threonine phosphate + H2O | - |
Mycoplasmopsis synoviae | [alpha-casein]-serine/threonine + phosphate | - |
? |
Subunits | Comment | Organism |
---|---|---|
? | x * 30000, about, recombinant His-tagged wild-type and mutant enzymes, SDS-PAGE | Mycoplasmopsis synoviae |
More | homology modeling and analysis of the three-dimensional structure of PrpC using the structure of Streptococcus agalactiae serine/threonine phosphatase as template, PDB ID 2PK0, overview. Comparison of PrpC structure with typical PP2C phosphatases | Mycoplasmopsis synoviae |
Synonyms | Comment | Organism |
---|---|---|
PPM phosphatase | - |
Mycoplasmopsis synoviae |
PrpC | - |
Mycoplasmopsis synoviae |
Ser/Thr phosphatase | - |
Mycoplasmopsis synoviae |
serine/threonine phosphatase | - |
Mycoplasmopsis synoviae |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
- |
Mycoplasmopsis synoviae |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
51.3 | 56.5 | thermal denaturation profile of His6-tagged wild-type apo-PrpC indicates a Tm of 56.5°C and 51.3°C for the detagged wild-type apoenzyme, while the mutant His6-tagged apo-PrpC R164A and D122A show a Tm of 54.8°C and 55.8°C, respectively | Mycoplasmopsis synoviae |
60 | - |
recombinant wild-type and mutant enzymes, stable up to | Mycoplasmopsis synoviae |
70 | - |
recombinant wild-type and mutant enzymes, almost inactivation | Mycoplasmopsis synoviae |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.09 | - |
RRA(pT)VA | pH 9.0, 30°C, recombinant wild-type enzyme | Mycoplasmopsis synoviae | |
1.3 | - |
[alpha-casein]-serine/threonine phosphate | pH 9.0, 30°C, recombinant wild-type enzyme | Mycoplasmopsis synoviae |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
9 | - |
- |
Mycoplasmopsis synoviae |
General Information | Comment | Organism |
---|---|---|
evolution | the enzyme belongs to the protein phosphatase 2C (PP2C) subfamily. Comparison of PrpC with typical PP2C phosphatases, overview | Mycoplasmopsis synoviae |
additional information | the conserved residues involved in PP2C third metal ion-binding site are also involved in PrpC catalysis, and the metal binding also contributes to the protein overall structural stability. Homology modeling and analysis of the three-dimensional structure of PrpC using the structure of Streptococcus agalactiae serine/threonine phosphatase as template, PDB ID 2PK0, overview | Mycoplasmopsis synoviae |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.292 | - |
RRA(pT)VA | pH 9.0, 30°C, recombinant wild-type enzyme | Mycoplasmopsis synoviae | |
56.5 | - |
[alpha-casein]-serine/threonine phosphate | pH 9.0, 30°C, recombinant wild-type enzyme | Mycoplasmopsis synoviae |