Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21(DE3) cells | Brucella melitensis |
Crystallization (Comment) | Organism |
---|---|
in complex with its product adenine, hanging drop vapor diffusion method, using 20% (w/v) PEG-1000, 0.1 M phosphate-citrate (pH 4.2) and 0.2 M Li2SO4 | Brucella melitensis |
Protein Variants | Comment | Organism |
---|---|---|
A15M | the mutant exhibits less than 16% catalytic efficiency compared to the wild type | Brucella melitensis |
D168A | inactive | Brucella melitensis |
D168N | inactive | Brucella melitensis |
E145A | inactive | Brucella melitensis |
E145Q | inactive | Brucella melitensis |
E18A | inactive | Brucella melitensis |
E18Q | inactive | Brucella melitensis |
M144A | the mutant exhibits less than 1% catalytic efficiency compared to the wild type | Brucella melitensis |
R164A | the variant displays catalytic efficiencies of less than 20% compared to the wild type | Brucella melitensis |
R85A | the mutant exhibits less than 8% catalytic efficiency compared to the wild type | Brucella melitensis |
R85H | the mutant exhibits less than 9% catalytic efficiency compared to the wild type | Brucella melitensis |
S167A | the mutant exhibits less than 6% catalytic efficiency compared to the wild type | Brucella melitensis |
V34A | the mutant exhibits less than 19% catalytic efficiency compared to the wild type | Brucella melitensis |
V34I | the mutant exhibits less than 22% catalytic efficiency compared to the wild type | Brucella melitensis |
W179A | the mutant exhibits less than 7% catalytic efficiency compared to the wild type | Brucella melitensis |
W179F | the mutant exhibits less than 3% catalytic efficiency compared to the wild type | Brucella melitensis |
Y134A | the mutant exhibits less than 1% catalytic efficiency compared to the wild type | Brucella melitensis |
Y134F | the mutant exhibits less than 1% catalytic efficiency compared to the wild type | Brucella melitensis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0022 | - |
S-methyl-5'-thioadenosine | wild type enzyme, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
0.0031 | - |
S-methyl-5'-thioadenosine | mutant enzyme A15M, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
0.004 | - |
S-adenosyl-L-homocysteine | wild type enzyme, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
0.0041 | - |
S-adenosyl-L-homocysteine | mutant enzyme A15M, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
0.0044 | - |
S-methyl-5'-thioadenosine | mutant enzyme S167A, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
0.0045 | - |
S-methyl-5'-thioadenosine | mutant enzyme V34I, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
0.0048 | - |
S-methyl-5'-thioadenosine | mutant enzyme R85H, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
0.0053 | - |
S-methyl-5'-thioadenosine | mutant enzyme V34A, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
0.0054 | - |
S-methyl-5'-thioadenosine | mutant enzyme R85A, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
0.0054 | - |
S-methyl-5'-thioadenosine | mutant enzyme W179A, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
0.0056 | - |
S-adenosyl-L-homocysteine | mutant enzyme S167A, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
0.0058 | - |
S-adenosyl-L-homocysteine | mutant enzyme V34I, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
0.0061 | - |
S-adenosyl-L-homocysteine | mutant enzyme R85H, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
0.0062 | - |
S-adenosyl-L-homocysteine | mutant enzyme V34A, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
0.0068 | - |
S-adenosyl-L-homocysteine | mutant enzyme R85A, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
0.0074 | - |
S-methyl-5'-thioadenosine | mutant enzyme W179F, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
0.0076 | - |
S-adenosyl-L-homocysteine | mutant enzyme W179A, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
0.008 | - |
S-methyl-5'-thioadenosine | mutant enzyme R164A, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
0.0098 | - |
S-adenosyl-L-homocysteine | mutant enzyme W179F, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
0.0133 | - |
S-adenosyl-L-homocysteine | mutant enzyme R164A, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
0.028 | - |
S-methyl-5'-thioadenosine | mutant enzyme Y134A, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
0.0301 | - |
S-methyl-5'-thioadenosine | mutant enzyme Y134F, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
0.0351 | - |
S-methyl-5'-thioadenosine | mutant enzyme M144A, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
0.0381 | - |
S-adenosyl-L-homocysteine | mutant enzyme Y134A, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
0.0452 | - |
S-adenosyl-L-homocysteine | mutant enzyme Y134F, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
0.0951 | - |
S-adenosyl-L-homocysteine | mutant enzyme M144A, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
25000 | - |
2 * 25000, His-tagged enzyme, calculated from amino acid sequence | Brucella melitensis |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
S-adenosyl-L-homocysteine + H2O | Brucella melitensis | - |
S-D-ribosyl-L-homocysteine + adenine | - |
ir | |
S-methyl-5'-thioadenosine + H2O | Brucella melitensis | - |
S-methyl-5-thio-D-ribose + adenine | - |
ir |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Brucella melitensis | Q8YBL1 | - |
- |
Purification (Comment) | Organism |
---|---|
ammonium sulfate precipitation, nickel affinity column chromatography, and Superdex 200 gel filtration | Brucella melitensis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
S-adenosyl-L-homocysteine + H2O | - |
Brucella melitensis | S-D-ribosyl-L-homocysteine + adenine | - |
ir | |
S-methyl-5'-thioadenosine + H2O | - |
Brucella melitensis | S-methyl-5-thio-D-ribose + adenine | - |
ir |
Subunits | Comment | Organism |
---|---|---|
homodimer | 2 * 25000, His-tagged enzyme, calculated from amino acid sequence | Brucella melitensis |
Synonyms | Comment | Organism |
---|---|---|
5'-methylthioadenosine/S-adenosylhomocysteine nucleosidase | - |
Brucella melitensis |
MTAN | - |
Brucella melitensis |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.15 | - |
S-adenosyl-L-homocysteine | mutant enzyme M144A, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
0.35 | - |
S-adenosyl-L-homocysteine | mutant enzyme Y134F, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
0.37 | - |
S-adenosyl-L-homocysteine | mutant enzyme W179F, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
0.39 | - |
S-adenosyl-L-homocysteine | mutant enzyme Y134A, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
0.51 | - |
S-methyl-5'-thioadenosine | mutant enzyme M144A, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
0.55 | - |
S-adenosyl-L-homocysteine | mutant enzyme S167A, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
0.58 | - |
S-methyl-5'-thioadenosine | mutant enzyme S167A, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
0.58 | - |
S-methyl-5'-thioadenosine | mutant enzyme W179F, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
0.69 | - |
S-methyl-5'-thioadenosine | mutant enzyme W179A, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
0.71 | - |
S-methyl-5'-thioadenosine | mutant enzyme Y134F, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
0.78 | - |
S-methyl-5'-thioadenosine | mutant enzyme Y134A, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
0.86 | - |
S-adenosyl-L-homocysteine | mutant enzyme W179A, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
0.92 | - |
S-adenosyl-L-homocysteine | mutant enzyme R85H, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
0.93 | - |
S-adenosyl-L-homocysteine | mutant enzyme R85A, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
1.1 | - |
S-adenosyl-L-homocysteine | mutant enzyme A15M, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
1.3 | - |
S-methyl-5'-thioadenosine | mutant enzyme R85H, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
1.4 | - |
S-methyl-5'-thioadenosine | mutant enzyme R85A, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
1.8 | - |
S-methyl-5'-thioadenosine | mutant enzyme A15M, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
2.1 | - |
S-adenosyl-L-homocysteine | mutant enzyme V34A, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
2.3 | - |
S-adenosyl-L-homocysteine | mutant enzyme V34I, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
2.5 | - |
S-methyl-5'-thioadenosine | mutant enzyme V34I, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
2.7 | - |
S-methyl-5'-thioadenosine | mutant enzyme V34A, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
4.7 | - |
S-adenosyl-L-homocysteine | mutant enzyme R164A, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
7.1 | - |
S-adenosyl-L-homocysteine | wild type enzyme, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
8 | - |
S-methyl-5'-thioadenosine | mutant enzyme R164A, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
8.8 | - |
S-methyl-5'-thioadenosine | wild type enzyme, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1.6 | - |
S-adenosyl-L-homocysteine | mutant enzyme M144A, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
7.8 | - |
S-adenosyl-L-homocysteine | mutant enzyme Y134F, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
10 | - |
S-adenosyl-L-homocysteine | mutant enzyme Y134A, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
15 | - |
S-methyl-5'-thioadenosine | mutant enzyme M144A, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
24 | - |
S-methyl-5'-thioadenosine | mutant enzyme Y134F, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
28 | - |
S-methyl-5'-thioadenosine | mutant enzyme Y134A, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
38 | - |
S-adenosyl-L-homocysteine | mutant enzyme W179F, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
79 | - |
S-methyl-5'-thioadenosine | mutant enzyme W179F, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
100 | - |
S-adenosyl-L-homocysteine | mutant enzyme S167A, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
110 | - |
S-adenosyl-L-homocysteine | mutant enzyme W179A, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
130 | - |
S-methyl-5'-thioadenosine | mutant enzyme S167A, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
130 | - |
S-methyl-5'-thioadenosine | mutant enzyme W179A, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
140 | - |
S-adenosyl-L-homocysteine | mutant enzyme R85A, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
150 | - |
S-adenosyl-L-homocysteine | mutant enzyme R85H, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
250 | - |
S-methyl-5'-thioadenosine | mutant enzyme R85A, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
270 | - |
S-adenosyl-L-homocysteine | mutant enzyme A15M, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
280 | - |
S-methyl-5'-thioadenosine | mutant enzyme R85H, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
330 | - |
S-adenosyl-L-homocysteine | mutant enzyme V34A, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
350 | - |
S-adenosyl-L-homocysteine | mutant enzyme R164A, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
390 | - |
S-adenosyl-L-homocysteine | mutant enzyme V34I, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
510 | - |
S-methyl-5'-thioadenosine | mutant enzyme V34A, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
560 | - |
S-methyl-5'-thioadenosine | mutant enzyme V34I, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
580 | - |
S-methyl-5'-thioadenosine | mutant enzyme A15M, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
600 | - |
S-methyl-5'-thioadenosine | mutant enzyme R164A, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
1800 | - |
S-adenosyl-L-homocysteine | wild type enzyme, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis | |
4000 | - |
S-methyl-5'-thioadenosine | wild type enzyme, in 100 mM HEPES (pH 7.5), at 25°C | Brucella melitensis |