EC Number | Cloned (Comment) | Organism |
---|---|---|
2.1.2.9 | expression of wild-type and of R42K, R42L and G41R/R42L mutant enzymes in Escherichia coli JM109 as His6-tagged proteins | Escherichia coli |
EC Number | Crystallization (Comment) | Organism |
---|---|---|
2.1.2.9 | - |
Escherichia coli |
EC Number | Protein Variants | Comment | Organism |
---|---|---|---|
2.1.2.9 | G41D | mutant enzyme with decreased activity | Escherichia coli |
2.1.2.9 | G41K | mutant enzyme with decreased activity | Escherichia coli |
2.1.2.9 | G41L | mutant enzyme with decreased activity | Escherichia coli |
2.1.2.9 | G41R | G41R/R42L double mutant with low activity, G41R: decreased activity | Escherichia coli |
2.1.2.9 | R38K | mutant enzyme with 2.2fold increased Km, no effect on kcat | Escherichia coli |
2.1.2.9 | R38L | mutant enzyme with up to 12fold lower specific activity, 6fold increased Km and 5fold decreased kcat | Escherichia coli |
2.1.2.9 | R42K | mutant enzyme with lower activity, 16fold increased Km | Escherichia coli |
2.1.2.9 | R42L | single mutant and G41R/R42L double mutant with low activity, R42L: 17fold increased Km and 79fold decreased kcat | Escherichia coli |
2.1.2.9 | R42Q | mutant enzyme with lower activity, 16-17fold increased Km | Escherichia coli |
EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
2.1.2.9 | additional information | - |
additional information | values for wild-type and mutant enzyme with wild-type and mutant initiator tRNAs | Escherichia coli | |
2.1.2.9 | 0.00054 | - |
L-methionyl-tRNAfMet | wild-type enzyme | Escherichia coli | |
2.1.2.9 | 0.00066 | - |
L-methionyl-tRNAfMet | G41R mutant enzyme | Escherichia coli | |
2.1.2.9 | 0.00102 | - |
L-methionyl-tRNAfMet | G41K mutant enzyme | Escherichia coli | |
2.1.2.9 | 0.00121 | - |
L-methionyl-tRNAfMet | R38K mutant enzyme | Escherichia coli | |
2.1.2.9 | 0.00162 | - |
L-methionyl-tRNAfMet | G41L mutant enzyme | Escherichia coli | |
2.1.2.9 | 0.003 | - |
L-methionyl-tRNAfMet | G41D mutant enzyme | Escherichia coli | |
2.1.2.9 | 0.00324 | - |
L-methionyl-tRNAfMet | R38L mutant enzyme | Escherichia coli | |
2.1.2.9 | 0.00834 | - |
L-methionyl-tRNAfMet | G41R/R42L double mutant enzyme | Escherichia coli | |
2.1.2.9 | 0.00866 | - |
L-methionyl-tRNAfMet | R42K mutant enzyme | Escherichia coli | |
2.1.2.9 | 0.009 | - |
L-methionyl-tRNAfMet | R42L mutant enzyme | Escherichia coli | |
2.1.2.9 | 0.0096 | - |
L-methionyl-tRNAfMet | R42Q mutant enzyme | Escherichia coli |
EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
2.1.2.9 | 10-formyltetrahydrofolate + L-methionyl-tRNAfMet + H2O | Escherichia coli | involved in initiation of protein biosynthesis | tetrahydrofolate + N-formylmethionyl-tRNAfMet | - |
? |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
2.1.2.9 | Escherichia coli | - |
- |
- |
EC Number | Purification (Comment) | Organism |
---|---|---|
2.1.2.9 | purification of wild-type and of R42K, R42L and G41R/R42L mutant recombinant enzymes, expressed in Escherichia coli JM109 | Escherichia coli |
EC Number | Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|---|
2.1.2.9 | additional information | - |
- |
Escherichia coli |
EC Number | Storage Stability | Organism |
---|---|---|
2.1.2.9 | -20°C, 20 mM imidazole, 10 mM 2-mercaptoethanol, 50% glycerol, 150 mM KCl, stable | Escherichia coli |
2.1.2.9 | -20°C, 20 mM imidazole, pH 7.5, 10 mM 2-mercaptoethanol, 50% glycerol, 150 mM KCl, 2-4 months, stable | Escherichia coli |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
2.1.2.9 | 10-formyltetrahydrofolate + L-methionyl-tRNAfMet | - |
Escherichia coli | tetrahydrofolate + N-formylmethionyl-tRNAfMet | - |
r | |
2.1.2.9 | 10-formyltetrahydrofolate + L-methionyl-tRNAfMet + H2O | specificity determinants at the end of the acceptor stem of tRNAfMet | Escherichia coli | tetrahydrofolate + N-formylmethionyl-tRNAfMet | - |
? | |
2.1.2.9 | 10-formyltetrahydrofolate + L-methionyl-tRNAfMet + H2O | invariant Arg-42 within the 16-amino acid insertion loop, residues 34-49, plays a very important role both in steps of substrate binding and in catalysis, Arg-42 makes functional contacts with the determinants in the acceptor stem of the initiator tRNA | Escherichia coli | tetrahydrofolate + N-formylmethionyl-tRNAfMet | - |
? | |
2.1.2.9 | 10-formyltetrahydrofolate + L-methionyl-tRNAfMet + H2O | specificity is restricted to methionyl-tRNAfMet | Escherichia coli | tetrahydrofolate + N-formylmethionyl-tRNAfMet | - |
? | |
2.1.2.9 | 10-formyltetrahydrofolate + L-methionyl-tRNAfMet + H2O | important role of a 16-amino acid insertion loop in enzyme for tRNA recognition | Escherichia coli | tetrahydrofolate + N-formylmethionyl-tRNAfMet | - |
? | |
2.1.2.9 | 10-formyltetrahydrofolate + L-methionyl-tRNAfMet + H2O | involved in initiation of protein biosynthesis | Escherichia coli | tetrahydrofolate + N-formylmethionyl-tRNAfMet | - |
? | |
2.1.2.9 | additional information | enzyme structure | Escherichia coli | ? | - |
? |
EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|---|
2.1.2.9 | 37 | - |
assay at | Escherichia coli |
EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
2.1.2.9 | additional information | - |
additional information | values for wild-type and mutant enzyme with wild-type and mutant initiator tRNAs | Escherichia coli | |
2.1.2.9 | 0.45 | - |
L-methionyl-tRNAfMet | G41R/R42L double mutant enzyme | Escherichia coli | |
2.1.2.9 | 0.483 | - |
L-methionyl-tRNAfMet | R42L mutant enzyme | Escherichia coli | |
2.1.2.9 | 3.1 | - |
L-methionyl-tRNAfMet | R42Q mutant enzyme | Escherichia coli | |
2.1.2.9 | 8.27 | - |
L-methionyl-tRNAfMet | R38L mutant enzyme | Escherichia coli | |
2.1.2.9 | 11.4 | - |
L-methionyl-tRNAfMet | G41R mutant enzyme | Escherichia coli | |
2.1.2.9 | 14.6 | - |
L-methionyl-tRNAfMet | G41K mutant enzyme | Escherichia coli | |
2.1.2.9 | 27.7 | - |
L-methionyl-tRNAfMet | R42K mutant enzyme | Escherichia coli | |
2.1.2.9 | 31.5 | - |
L-methionyl-tRNAfMet | G41L mutant enzyme | Escherichia coli | |
2.1.2.9 | 33 | - |
L-methionyl-tRNAfMet | G41D mutant enzyme | Escherichia coli | |
2.1.2.9 | 39.1 | - |
L-methionyl-tRNAfMet | wild-type enzyme | Escherichia coli | |
2.1.2.9 | 46.3 | - |
L-methionyl-tRNAfMet | R38K mutant enzyme | Escherichia coli |