Cloned (Comment) | Organism |
---|---|
cloning of the fmt gene and expression of wild-type and mutant genes in Escherichia coli BL21(DE3) as His6-tagged proteins | Haemophilus influenzae |
fmt gene encoding enzyme is cloned | Thermus thermophilus |
fmt gene encoding enzyme is cloned | Pseudomonas aeruginosa |
fmt gene encoding enzyme is cloned | Rickettsia prowazekii |
Protein Variants | Comment | Organism |
---|---|---|
D145A | mutant enzyme with drastically reduced activity | Haemophilus influenzae |
D145E | mutant enzyme with 881fold reduced formylation rate, kinetic parameters | Haemophilus influenzae |
D145N | mutant enzyme with 4031fold reduced formylation rate, kinetic parameters | Haemophilus influenzae |
H109A | mutant enzyme with 4677fold reduced formylation rate, kinetic parameters | Haemophilus influenzae |
H109K | inactive mutant enzyme | Haemophilus influenzae |
H109R | mutant enzyme with 7172fold reduced formylation rate, kinetic parameters | Haemophilus influenzae |
K37A | mutant enzyme with 3.5fold reduced formylation rate, kinetic parameters | Haemophilus influenzae |
additional information | mutations of Asp-145 and Arg-41 reduce the affinity for initiator tRNA, mutations of Asn-107 and His-109 affect catalysis but not tRNA binding | Haemophilus influenzae |
N107A | mutant enzyme with 48fold reduced formylation rate, kinetic parameters | Haemophilus influenzae |
R41A | mutant enzyme with 470fold reduced formylation rate, kinetic parameters | Haemophilus influenzae |
R41K | mutant enzyme with 140fold reduced formylation rate, kinetic parameters | Haemophilus influenzae |
V31A | mutant enzyme with 1.7fold reduced formylation rate | Haemophilus influenzae |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | values for several mutant enzymes | Haemophilus influenzae | |
0.00112 | - |
E.coli L-methionyl-tRNAfMet | - |
Haemophilus influenzae |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Haemophilus influenzae | - |
- |
- |
Pseudomonas aeruginosa | - |
- |
- |
Rickettsia prowazekii | - |
- |
- |
Thermus thermophilus | - |
- |
- |
Purification (Comment) | Organism |
---|---|
purification of His6-tagged wild-type and mutant recombinant enzymes, expressed in Escherichia coli BL21(DE3) | Haemophilus influenzae |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
- |
Haemophilus influenzae |
1.23 | - |
- |
Haemophilus influenzae |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
10-formyltetrahydrofolate + E. coli L-methionyl-tRNAfMet | - |
Haemophilus influenzae | tetrahydrofolate + N-formylmethionyl-tRNAfMet | - |
r | |
10-formyltetrahydrofolate + E.coli L-methionyl-tRNAfMet | - |
Haemophilus influenzae | tetrahydrofolate + E.coli N-formylmethionyl-tRNAfMet | - |
r | |
10-formyltetrahydrofolate + L-methionyl-tRNAfMet + H2O | formylates methionyl-tRNAfMet from E. coli | Haemophilus influenzae | tetrahydrofolate + N-formylmethionyl-tRNAfMet | - |
? | |
10-formyltetrahydrofolate + L-methionyl-tRNAfMet + H2O | Asn-107, His-109 and Asp-145 are catalytic residues, Arg-41 is involved in tRNA recognition | Haemophilus influenzae | tetrahydrofolate + N-formylmethionyl-tRNAfMet | - |
? |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Haemophilus influenzae |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | values for several mutant enzymes | Haemophilus influenzae | |
11.3 | - |
E. coli L-methionyl-tRNAfMet | - |
Haemophilus influenzae |