Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21(DE3) cells | Brucella melitensis |
Crystallization (Comment) | Organism |
---|---|
in complex with 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 shows 14.5% catalytic efficiency compared to the wild type enzyme | Brucella melitensis |
D168A | the mutation completely abolishes activity | Brucella melitensis |
D168N | the mutation completely abolishes activity | Brucella melitensis |
E145A | the mutation completely abolishes activity | Brucella melitensis |
E145Q | the mutation completely abolishes activity | Brucella melitensis |
E18A | the mutation completely abolishes activity | Brucella melitensis |
E18Q | the mutation completely abolishes activity | Brucella melitensis |
M144A | the mutant shows 0.4% catalytic efficiency compared to the wild type enzyme | Brucella melitensis |
R164A | the mutant shows 15% catalytic efficiency compared to the wild type enzyme | Brucella melitensis |
R85A | the mutant shows 6.3% catalytic efficiency compared to the wild type enzyme | Brucella melitensis |
R85H | the mutant shows 7.1% catalytic efficiency compared to the wild type enzyme | Brucella melitensis |
S167A | the mutant shows 3.3% catalytic efficiency compared to the wild type enzyme | Brucella melitensis |
V34A | the mutant shows 12.5% catalytic efficiency compared to the wild type enzyme | Brucella melitensis |
V34I | the mutant shows 13.9% catalytic efficiency compared to the wild type enzyme | Brucella melitensis |
W179A | the mutant shows 3.3% catalytic efficiency compared to the wild type enzyme | Brucella melitensis |
W179F | the mutant shows 2.0% catalytic efficiency compared to the wild type enzyme | Brucella melitensis |
Y134A | the mutant shows 8.1% catalytic efficiency compared to the wild type enzyme | Brucella melitensis |
Y134F | the mutant shows 0.6% catalytic efficiency compared to the wild type enzyme | Brucella melitensis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0022 | - |
S-methyl-5'-thioadenosine | wild type enzyme, at pH 7.0 and 25°C | Brucella melitensis | |
0.0031 | - |
S-methyl-5'-thioadenosine | mutant enzyme A15M, at pH 7.0 and 25°C | Brucella melitensis | |
0.0044 | - |
S-methyl-5'-thioadenosine | mutant enzyme S167A, at pH 7.0 and 25°C | Brucella melitensis | |
0.0045 | - |
S-methyl-5'-thioadenosine | mutant enzyme V34I, at pH 7.0 and 25°C | Brucella melitensis | |
0.0048 | - |
S-methyl-5'-thioadenosine | mutant enzyme R85H, at pH 7.0 and 25°C | Brucella melitensis | |
0.0053 | - |
S-methyl-5'-thioadenosine | mutant enzyme V34A, at pH 7.0 and 25°C | Brucella melitensis | |
0.0054 | - |
S-methyl-5'-thioadenosine | mutant enzyme R85A, at pH 7.0 and 25°C | Brucella melitensis | |
0.0054 | - |
S-methyl-5'-thioadenosine | mutant enzyme W179A, at pH 7.0 and 25°C | Brucella melitensis | |
0.0074 | - |
S-methyl-5'-thioadenosine | mutant enzyme W179F, at pH 7.0 and 25°C | Brucella melitensis | |
0.028 | - |
S-methyl-5'-thioadenosine | mutant enzyme Y134A, at pH 7.0 and 25°C | Brucella melitensis | |
0.0301 | - |
S-methyl-5'-thioadenosine | mutant enzyme Y134F, at pH 7.0 and 25°C | Brucella melitensis | |
0.0351 | - |
S-methyl-5'-thioadenosine | mutant enzyme M144A, at pH 7.0 and 25°C | Brucella melitensis | |
8 | - |
S-methyl-5'-thioadenosine | mutant enzyme R164A, at pH 7.0 and 25°C | Brucella melitensis |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
S-methyl-5'-thioadenosine + phosphate | Brucella melitensis | - |
adenine + S-methyl-5-thio-alpha-D-ribose 1-phosphate | - |
ir |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Brucella melitensis | Q8YBL1 | - |
- |
Purification (Comment) | Organism |
---|---|
nickel affinity column chromatography, and Superdex 200 gel filtration | Brucella melitensis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
S-methyl-5'-thioadenosine + phosphate | - |
Brucella melitensis | adenine + S-methyl-5-thio-alpha-D-ribose 1-phosphate | - |
ir |
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 |
---|---|---|---|---|---|
0.51 | - |
S-methyl-5'-thioadenosine | mutant enzyme M144A, at pH 7.0 and 25°C | Brucella melitensis | |
0.58 | - |
S-methyl-5'-thioadenosine | mutant enzyme S167A, at pH 7.0 and 25°C | Brucella melitensis | |
0.58 | - |
S-methyl-5'-thioadenosine | mutant enzyme W179F, at pH 7.0 and 25°C | Brucella melitensis | |
0.69 | - |
S-methyl-5'-thioadenosine | mutant enzyme W179A, at pH 7.0 and 25°C | Brucella melitensis | |
0.71 | - |
S-methyl-5'-thioadenosine | mutant enzyme Y134F, at pH 7.0 and 25°C | Brucella melitensis | |
0.78 | - |
S-methyl-5'-thioadenosine | mutant enzyme Y134A, at pH 7.0 and 25°C | Brucella melitensis | |
1.3 | - |
S-methyl-5'-thioadenosine | mutant enzyme R85H, at pH 7.0 and 25°C | Brucella melitensis | |
1.4 | - |
S-methyl-5'-thioadenosine | mutant enzyme R85A, at pH 7.0 and 25°C | Brucella melitensis | |
1.8 | - |
S-methyl-5'-thioadenosine | mutant enzyme A15M, at pH 7.0 and 25°C | Brucella melitensis | |
2.5 | - |
S-methyl-5'-thioadenosine | mutant enzyme V34I, at pH 7.0 and 25°C | Brucella melitensis | |
2.7 | - |
S-methyl-5'-thioadenosine | mutant enzyme V34A, at pH 7.0 and 25°C | Brucella melitensis | |
8 | - |
S-methyl-5'-thioadenosine | mutant enzyme R164A, at pH 7.0 and 25°C | Brucella melitensis | |
8.8 | - |
S-methyl-5'-thioadenosine | wild type enzyme, at pH 7.0 and 25°C | Brucella melitensis |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
15 | - |
S-methyl-5'-thioadenosine | mutant enzyme M144A, at pH 7.0 and 25°C | Brucella melitensis | |
24 | - |
S-methyl-5'-thioadenosine | mutant enzyme Y134F, at pH 7.0 and 25°C | Brucella melitensis | |
28 | - |
S-methyl-5'-thioadenosine | mutant enzyme Y134A, at pH 7.0 and 25°C | Brucella melitensis | |
79 | - |
S-methyl-5'-thioadenosine | mutant enzyme W179F, at pH 7.0 and 25°C | Brucella melitensis | |
130 | - |
S-methyl-5'-thioadenosine | mutant enzyme S167A, at pH 7.0 and 25°C | Brucella melitensis | |
130 | - |
S-methyl-5'-thioadenosine | mutant enzyme W179A, at pH 7.0 and 25°C | Brucella melitensis | |
250 | - |
S-methyl-5'-thioadenosine | mutant enzyme R85A, at pH 7.0 and 25°C | Brucella melitensis | |
280 | - |
S-methyl-5'-thioadenosine | mutant enzyme R85H, at pH 7.0 and 25°C | Brucella melitensis | |
510 | - |
S-methyl-5'-thioadenosine | mutant enzyme V34A, at pH 7.0 and 25°C | Brucella melitensis | |
560 | - |
S-methyl-5'-thioadenosine | mutant enzyme V34I, at pH 7.0 and 25°C | Brucella melitensis | |
580 | - |
S-methyl-5'-thioadenosine | mutant enzyme A15M, at pH 7.0 and 25°C | Brucella melitensis | |
600 | - |
S-methyl-5'-thioadenosine | mutant enzyme R164A, at pH 7.0 and 25°C | Brucella melitensis | |
4000 | - |
S-methyl-5'-thioadenosine | wild type enzyme, at pH 7.0 and 25°C | Brucella melitensis |