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Literature summary for 2.3.2.10 extracted from
- Kinetic and mechanistic characterization of recombinant Lactobacillus viridescens FemX (UDP-N-acetylmuramoyl pentapeptide-lysine N6-alanyltransferase) (2003), J. Biol. Chem., 278, 22861-22867.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| expression of wild-type and mutants as His-tagged proteins in Escherichia coli, in vitro transcription of tRNAs | Weissella viridescens |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| D109N | site-directed mutagenesis, loss of 99% activity compared to the wild-type | Weissella viridescens |
| E205Q | site-directed mutagenesis, unaltered activity compared to the wild-type | Weissella viridescens |
| E215Q | site-directed mutagenesis, loss of 15% activity compared to the wild-type | Weissella viridescens |
| E316Q | site-directed mutagenesis, loss of 60% activity compared to the wild-type | Weissella viridescens |
| E320Q | site-directed mutagenesis, loss of 96% activity compared to the wild-type | Weissella viridescens |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| Ser-tRNA | versus UDP-acetylmuramoylpentapeptide | Weissella viridescens |  |
| UDP-N-acetylmuramoyl-L-alanyl-D-glutamyl-L-lysyl-D-alanyl-D-alanine | product inhibition | Weissella viridescens | |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.015 | - |
L-alanyl-tRNA(Ala) | Ala-tRNAAla from E. coli, recombinant wild-type | Weissella viridescens | |
| 0.042 | - |
UDP-acetylmuramoylpentapeptide | recombinant wild-type | Weissella viridescens | |
| 0.054 | - |
L-alanyl-tRNA(Ala) | Ala-tRNAAla from E. coli, recombinant mutant E320Q | Weissella viridescens | |
| 0.061 | - |
L-alanyl-tRNA(Ala) | Ala-tRNAAla from E. coli, recombinant mutant D109N | Weissella viridescens | |
| 0.067 | - |
UDP-acetylmuramoylpentapeptide | recombinant mutant E320Q | Weissella viridescens | |
| 0.165 | - |
UDP-acetylmuramoylpentapeptide | recombinant mutant D109N | Weissella viridescens | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Weissella viridescens | - |
femX gene | - |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant His-tagged wild-type and mutants from E. coli | Weissella viridescens |
Reaction
| Reaction | Comment | Organism | Reaction ID |
|---|---|---|---|
| L-alanyl-tRNAAla + UDP-N-acetyl-alpha-D-muramoyl-L-alanyl-D-glutamyl-L-lysyl-D-alanyl-D-alanine = tRNAAla + UDP-N-acetyl-alpha-D-muramoyl-L-alanyl-D-glutamyl-N6-(L-alanyl)-L-lysyl-D-alanyl-D-alanine | transfer of L-Ala to epsilon-amino group of L-lysine of UDP-UDP-acetylmuramoylpentapeptide in an ordered sequential mechanism | Weissella viridescens | |
| L-alanyl-tRNAAla + UDP-N-acetyl-alpha-D-muramoyl-L-alanyl-D-glutamyl-L-lysyl-D-alanyl-D-alanine = tRNAAla + UDP-N-acetyl-alpha-D-muramoyl-L-alanyl-D-glutamyl-N6-(L-alanyl)-L-lysyl-D-alanyl-D-alanine | catalysis via a ternary complex | Weissella viridescens |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| L-alanyl-tRNA(Ala) + UDP-N-acetylmuramoyl-pentapeptide containing L-lysine | - |
Weissella viridescens | ? | - |
r | |
| L-alanyl-tRNAAla + UDP-N-acetylmuramoyl-L-alanyl-D-glutamyl-L-lysyl-D-alanyl-D-alanine | - |
Weissella viridescens | tRNAAla + UDP-N-acetylmuramoyl-L-alanyl-D-glutamyl-N6-(L-alanyl)-L-lysyl-D-alanyl-D-alanine | - |
? | |
| L-alanyl-tRNASer + UDP-N-acetylmuramoyl-L-alanyl-D-glutamyl-L-lysyl-D-alanyl-D-alanine | - |
Weissella viridescens | tRNASer + UDP-N-acetylmuramoyl-L-alanyl-D-glutamyl-N6-(L-alanyl)-L-lysyl-D-alanyl-D-alanine | - |
? | |
| L-seryl-tRNASer + UDP-N-acetylmuramoyl-L-alanyl-D-glutamyl-L-lysyl-D-alanyl-D-alanine | - |
Weissella viridescens | tRNASer + UDP-N-acetylmuramoyl-L-alanyl-D-glutamyl-N6-(L-seryl)-L-lysyl-D-alanyl-D-alanine | - |
? | |
| additional information | also utilizes UDP-N-acetylmuramoylpentapeptide containing L-ornithine as substrate | Weissella viridescens | ? | - |
? | |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 25 | - |
assay at | Weissella viridescens |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.183 | - |
L-alanyl-tRNAAla | recombinant mutant D109N | Weissella viridescens | |
| 0.917 | - |
L-alanyl-tRNAAla | recombinant mutant E320Q | Weissella viridescens | |
| 11 | - |
L-alanyl-tRNAAla | recombinant wild-type | Weissella viridescens | |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| additional information | - |
broad maximum | Weissella viridescens |
pH Range
| pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|
| additional information | - |
pK values of 5.5 and 9.3 | Weissella viridescens |
| 5.5 | 9.1 | - |
Weissella viridescens |
Ki Value [mM]
| Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
|---|---|---|---|---|---|
| additional information | - |
additional information | - |
Weissella viridescens | |
| 0.048 | - |
UDP-N-acetylmuramoyl-L-alanyl-D-glutamyl-L-lysyl-D-alanyl-D-alanine | product inhibition | Weissella viridescens | |
| 0.248 | - |
Ser-tRNA | versus UDP-acetylmuramoylpentapeptide | Weissella viridescens | |
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