Crystallization (Comment) | Organism |
---|---|
purified enzyme, oil microbatch method, mixing of 0.001 ml of 40 mg/ml protein solution in 50 mM MES-NaOH, pH 6.5, with 0.001 ml of precipitant solution composed of 250 mM NaNO3, 50 mM CaCl2, and 50 mM MES-NaOH, pH 6.5, to form Pr3+-derivatized crystals, the precipitant solution containing additionally 25 mM PrCl3 is used, X-ray diffraction structure determination and analysis at 1.45 A resolution | Parengyodontium album |
Protein Variants | Comment | Organism |
---|---|---|
molecular biology | proteinase K is widely used in molecular biology for its broad substrate specificity, wide pH stability, and high hydrolysis activity. Aminolysis by proteinase K is also attractive for chemoenzymatic peptide synthesis | Parengyodontium album |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Ca2+ | 1 mM CaCl2, the denaturation temperature of proteinase K derivatized with praseodymium (Pr) ions is 16.2°C, which is 5.9°C higher than those of metal-free and Ca2+-bound proteinase K, respectively. Isothermal titration calorimetry (ITC) measurements demonstrate that Pr-ion binding to proteinase K shows endothermic peaks, whereas Ca2+-ion binding shows exothermic peaks, indicating that the binding mode of Pr ions is different from that of Ca2+ ions, even though the crystal structures of proteinase K with Pr and Ca2+ ions are identical | Parengyodontium album | |
dysprosium | 1 mM DyCl3, differential scanning calorimetry analysis bound to the enzyme | Parengyodontium album | |
europium | 1 mM EuCl33, differential scanning calorimetry analysis bound to the enzyme | Parengyodontium album | |
gadolinium | 1 mM GdCl33, differential scanning calorimetry analysis bound to the enzyme | Parengyodontium album | |
holmium | 1 mM HoCl3, differential scanning calorimetry analysis bound to the enzyme | Parengyodontium album | |
lanthanum | 1 mM La(NO3)3, differential scanning calorimetry analysis bound to the enzyme | Parengyodontium album | |
lutetium | 1 mM LuCl3, differential scanning calorimetry analysis bound to the enzyme | Parengyodontium album | |
additional information | differential scanning calorimetry curves for proteinase K derivatized with heavy atoms, showing the correlation between atomic number and denaturation temperature, overview | Parengyodontium album | |
neodymium | 1 mM NdCl3, differential scanning calorimetry analysis bound to the enzyme | Parengyodontium album | |
Pr3+ | 1 mM PrCl3, the denaturation temperature of proteinase K derivatized with praseodymium (Pr) ions is 16.2°C, which is 5.9°C higher than those of metal-free and Ca2+-bound proteinase K, respectively. Isothermal titration calorimetry (ITC) measurements demonstrate that Pr-ion binding to proteinase K shows endothermic peaks, whereas Ca2+-ion binding shows exothermic peaks, indicating that the binding mode of Pr ions is different from that of Ca2+ ions, even though the crystal structures of proteinase K with Pr and Ca2+ ions are identical. Hydrolytic activity of Pr-derivatized proteinase K shows that the hydrolytic activity is 46fold higher at 70°C using synthetic nitroanilide substrate and 9 and 76fold higher at 70°C and 80°C using fluorescein isothiocyanate-labeled casein, respectively, in comparison with the native proteinase K. Furthermore, based on the yield of chemoenzymatic peptide syntheses, the aminolysis activity of Pr-derivatized proteinase K is 3.5 and 9.5fold higher than that of the native proteinase K at 50°C and 60°C, respectively. Analysis of the mechanism by which Pr ions enhance the thermal stability of proteinase K, overview | Parengyodontium album | |
samarium | 1 mM SmCl3, differential scanning calorimetry analysis bound to the enzyme | Parengyodontium album | |
ytterbium | 1 mM YbCl3, differential scanning calorimetry analysis bound to the enzyme | Parengyodontium album |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Parengyodontium album | P06873 | i.e. Tritirachium album or Engyodontium album | - |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
commercial preparation | - |
Parengyodontium album | - |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | the enzyme has a broad substrate specificity. Evaluation of aminolytic activity by polymerization of glutamic acid diethyl ester oligo(glutamic acid ethyl ester) | Parengyodontium album | ? | - |
? | |
N-succinyl-L-Phe-4-nitroanilide + H2O | - |
Parengyodontium album | N-succinyl-L-Phe + 4-nitroaniline | - |
? |
Synonyms | Comment | Organism |
---|---|---|
Proteinase K | - |
Parengyodontium album |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
additional information | - |
maximum degree of polymerization is achieved at 60°C using Pr-derivatized proteinase K, and at 50°C using the native proteinase K with Ca2+ ions. The yield of oligo(glutamic acid ethyl ester) is highest at 30°C using both the native and Pr-derivatized proteinase K | Parengyodontium album |
70 | - |
hydrolytic activity | Parengyodontium album |
Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | 80 | activity range, profile overview | Parengyodontium album |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
16.2 | 80 | the denaturation temperature of proteinase K derivatized with praseodymium (Pr) ions is 16.2°C, which is 5.9°C higher than those of metal-free and Ca2+-bound proteinase K, respectively. Isothermal titration calorimetry (ITC) measurements demonstrate that Pr-ion binding to proteinase K shows endothermic peaks, whereas Ca2+-ion binding shows exothermic peaks, indicating that the binding mode of Pr ions is different from that of Ca2+ ions, even though the crystal structures of proteinase K with Pr and Ca2+ ions are identical. Hydrolytic activity of Pr-derivatized proteinase K shows that the hydrolytic activity is 46fold higher at 70°C using synthetic nitroanilide substrate and 9 and 76fold higher at 70°C and 80°C using fluorescein isothiocyanate-labeled casein, respectively, in comparison with the native proteinase K. Furthermore, based on the yield of chemoenzymatic peptide syntheses, the aminolysis activity of Pr-derivatized proteinase K is 3.5 and 9.5fold higher than that of the native proteinase K at 50°C and 60°C, respectively | Parengyodontium album |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6.5 | - |
assay at | Parengyodontium album |