Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli strain BL21(DE3) | Mycobacterium tuberculosis |
expressed in Escherichia coli strain BL21(DE3) | Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
H98A | the mutation suppresses the 2',3'-cAMP phosphodiesterase activity of Rv0805 without adversely affecting hydrolysis of bis-p-nitrophenyl phosphate | Mycobacterium tuberculosis |
H98A | the mutation suppresses the 2',3'-cAMP phosphodiesterase activity of Rv0805 without adversely affecting hydrolysis of bis-p-nitrophenyl phosphate | Escherichia coli |
H98D | the mutation suppresses the 2',3'-cAMP phosphodiesterase activity of Rv0805 without adversely affecting hydrolysis of bis-p-nitrophenyl phosphate | Mycobacterium tuberculosis |
H98D | the mutation suppresses the 2',3'-cAMP phosphodiesterase activity of Rv0805 without adversely affecting hydrolysis of bis-p-nitrophenyl phosphate | Escherichia coli |
H98N | the mutation suppresses the 2',3'-cAMP phosphodiesterase specific activity to one-fifth the level of wild type Rv0805 | Mycobacterium tuberculosis |
H98N | the mutation suppresses the 2',3'-cAMP phosphodiesterase specific activity to one-fifth the level of wild type Rv0805 | Escherichia coli |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
phosphate | - |
Escherichia coli |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.9 | - |
bis-p-nitrophenyl phosphate | wild type enzyme, in Tris-HCl buffer (pH 8.5) in the presence of 0.5 mM MnCl2 | Mycobacterium tuberculosis | |
1.6 | - |
cyclic 2',3'-AMP | wild type enzyme, in Tris-HCl buffer (pH 8.5) in the presence of 0.5 mM MnCl2 | Mycobacterium tuberculosis | |
1.7 | - |
p-nitrophenyl phosphate | wild type enzyme, in Tris-HCl buffer (pH 8.5) in the presence of 0.5 mM MnCl2 | Mycobacterium tuberculosis | |
6 | - |
p-nitrophenyl phosphate | in Tris-HCl buffer (pH 8.5) in the presence of 5 mM MnCl2 | Escherichia coli | |
9.3 | - |
bis-p-nitrophenyl phosphate | in Tris-HCl buffer (pH 8.5) in the presence of 0.5 mM MnCl2 | Escherichia coli | |
35 | - |
cyclic 2',3'-AMP | in Tris-HCl buffer (pH 8.5) in the presence of 0.5 mM MnCl2 | Escherichia coli |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mn2+ | dependent on | Mycobacterium tuberculosis | |
Mn2+ | dependent on | Escherichia coli |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | P67095 | - |
- |
Mycobacterium tuberculosis | - |
- |
- |
Purification (Comment) | Organism |
---|---|
Ni-NTA agarose column chromatography | Mycobacterium tuberculosis |
Ni-NTA agarose column chromatography | Escherichia coli |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
bis-p-nitrophenyl phosphate + H2O | - |
Mycobacterium tuberculosis | p-nitrophenol + p-nitrophenyl phosphate | - |
? | |
bis-p-nitrophenyl phosphate + H2O | - |
Escherichia coli | p-nitrophenol + p-nitrophenyl phosphate | - |
? | |
cyclic 2',3'-AMP + H2O | Rv0805 is 150fold more active in hydrolyzing 2',3'-cAMP than 3',5'-cAMP, the enzyme hydrolyzes either P-O2' or P-O3' bonds to yield a mixture of 3'-AMP and 2'-AMP products, with a bias toward 3'-AMP | Mycobacterium tuberculosis | 3'-AMP | - |
? | |
cyclic 2',3'-AMP + H2O | YfcE-C74H cleaves exclusively the P-O2' bond of 2',3'-cAMP | Escherichia coli | 3'-AMP | - |
? | |
cyclic 2',3'-GMP + H2O | - |
Mycobacterium tuberculosis | 3'-GMP | - |
? | |
cyclic 2',3'-GMP + H2O | - |
Escherichia coli | 3'-GMP | - |
? | |
cyclic 3',5'-AMP + H2O | weak activity | Escherichia coli | 5'-AMP | - |
? | |
cyclic 3',5'-AMP + H2O | weak activity, Rv0805 is 150fold more active in hydrolyzing 2',3'-cAMP than 3',5'-cAMP | Mycobacterium tuberculosis | 5'-AMP | - |
? | |
cyclic 3',5'-GMP + H2O | weak activity | Mycobacterium tuberculosis | 5'-GMP | - |
? | |
cyclic 3',5'-GMP + H2O | weak activity | Escherichia coli | 5'-GMP | - |
? | |
cyclic 3',5'-UMP + H2O | weak activity | Mycobacterium tuberculosis | 5'-UMP | - |
? | |
cyclic 3',5'-UMP + H2O | weak activity | Escherichia coli | 5'-UMP | - |
? | |
additional information | the enzyme does not have a preference for 2',3'-cAMP versus 2',3'-cGMP | Mycobacterium tuberculosis | ? | - |
? | |
additional information | YfcE-C74H has no detectable ability to hydrolyze phosphomonoester substrates 5'-AMP, 3'-AMP, or 2'-AMP | Escherichia coli | ? | - |
? | |
p-nitrophenyl phosphate + H2O | - |
Escherichia coli | p-nitrophenol + phosphate | - |
? | |
p-nitrophenyl phosphate + H2O | weak activity | Mycobacterium tuberculosis | p-nitrophenol + phosphate | - |
? |
Synonyms | Comment | Organism |
---|---|---|
2,3-cyclic nucleotide phosphodiesterase | - |
Mycobacterium tuberculosis |
Rv0805 | functions as a 3',5'-cyclic nucleotide phosphodiesterase but has also 2',3'-cyclic nucleotide phosphodiesterase function | Mycobacterium tuberculosis |
YfcE-C74H | the mutant enzyme is strictly specific for 2',3'-cNMP, the C74H mutation uniquely confers a gain of function in hydrolysis of 2',3'-cAMP | Escherichia coli |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.55 | - |
p-nitrophenyl phosphate | wild type enzyme, in Tris-HCl buffer (pH 8.5) in the presence of 0.5 mM MnCl2 | Mycobacterium tuberculosis | |
2.8 | - |
cyclic 2',3'-AMP | wild type enzyme, in Tris-HCl buffer (pH 8.5) in the presence of 0.5 mM MnCl2 | Mycobacterium tuberculosis | |
2.95 | - |
cyclic 2',3'-AMP | in Tris-HCl buffer (pH 8.5) in the presence of 0.5 mM MnCl2 | Escherichia coli | |
3 | - |
p-nitrophenyl phosphate | in Tris-HCl buffer (pH 8.5) in the presence of 0.5 mM MnCl2 | Escherichia coli | |
12.4 | - |
bis-p-nitrophenyl phosphate | wild type enzyme, in Tris-HCl buffer (pH 8.5) in the presence of 0.5 mM MnCl2 | Mycobacterium tuberculosis | |
72 | - |
bis-p-nitrophenyl phosphate | in Tris-HCl buffer (pH 8.5) in the presence of 0.5 mM MnCl2 | Escherichia coli |
IC50 Value | IC50 Value Maximum | Comment | Organism | Inhibitor | Structure |
---|---|---|---|---|---|
2.5 | - |
- |
Escherichia coli | phosphate |