Crystallization (Comment) | Organism |
---|---|
structural comparison, molecular dynamics simulations, and site-directed mutagenesis suggest the positioning of the iron, ectoine, and 2-oxoglutarate ligands in close proximity to each other and with a spatial orientation that will allow the region-selective and stereo-specific hydroxylation of (4S)-ectoine to (4S,5S)-5-hydroxyectoine | Virgibacillus salexigens |
Protein Variants | Comment | Organism |
---|---|---|
A163C | residue is not involved in ligand binding | Virgibacillus salexigens |
A163C/S244C | residues are not involved in ligand binding | Virgibacillus salexigens |
D148A | loss of activity. Residue is involved in binding of Fe2+ | Virgibacillus salexigens |
D148E | loss of activity. Residue is involved in binding of Fe2+ | Virgibacillus salexigens |
F143A | loss of activity. Residue is involved in binding of 2-oxoglutarate | Virgibacillus salexigens |
F143W | loss of activity. Residue is involved in binding of 2-oxoglutarate | Virgibacillus salexigens |
F143Y | loss of activity. Residue is involved in binding of 2-oxoglutarate | Virgibacillus salexigens |
F242A | loss of activity. Residue is involved in binding of ectoine | Virgibacillus salexigens |
F242W | loss of activity. Residue is involved in binding of ectoine | Virgibacillus salexigens |
F242Y | 3fold increase in Km value | Virgibacillus salexigens |
F263A | loss of activity. Residue is involved in binding of ectoine | Virgibacillus salexigens |
F263W | loss of activity. Residue is involved in binding of ectoine | Virgibacillus salexigens |
F263Y | 30% increase in Km value | Virgibacillus salexigens |
F95A | loss of activity. Residue is involved in binding of 2-oxoglutarate | Virgibacillus salexigens |
H146A | loss of activity. Residue is involved in binding of Fe2+ | Virgibacillus salexigens |
H248A | loss of activity. Residue is involved in binding of Fe2+ | Virgibacillus salexigens |
N133A | loss of activity. Residue is involved in binding of 2-oxoglutarate | Virgibacillus salexigens |
N261A | residue is not involved in ligand binding | Virgibacillus salexigens |
P198A | activity similar to wild-type, residue of loop region | Virgibacillus salexigens |
Q129A | loss of activity. Residue is involved in binding of ectoine | Virgibacillus salexigens |
R131A | loss of activity. Residue is involved in binding of 2-oxoglutarate | Virgibacillus salexigens |
R259A | loss of activity. Residue is involved in binding of 2-oxoglutarate | Virgibacillus salexigens |
R259H | loss of activity. Residue is involved in binding of 2-oxoglutarate | Virgibacillus salexigens |
R259K | loss of activity. Residue is involved in binding of 2-oxoglutarate | Virgibacillus salexigens |
R259Q | loss of activity. Residue is involved in binding of 2-oxoglutarate | Virgibacillus salexigens |
S165A | 3fold increase in Km value | Virgibacillus salexigens |
S167A | residue is not involved in ligand binding | Virgibacillus salexigens |
S205A | activity similar to wild-type, residue of loop region | Virgibacillus salexigens |
S244C | residue is not involved in ligand binding | Virgibacillus salexigens |
S250A | loss of activity. Residue is involved in binding of 2-oxoglutarate | Virgibacillus salexigens |
V265A | residue is not involved in ligand binding | Virgibacillus salexigens |
V265L | residue is not involved in ligand binding | Virgibacillus salexigens |
V265T | residue is not involved in ligand binding | Virgibacillus salexigens |
W152A | loss of activity. Residue is involved in binding of ectoine | Virgibacillus salexigens |
W152F | loss of activity. Residue is involved in binding of ectoine | Virgibacillus salexigens |
W152Y | loss of activity. Residue is involved in binding of ectoine | Virgibacillus salexigens |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
5.9 | - |
ectoine | wild-type, pH 7.5, 32°C | Virgibacillus salexigens | |
6.8 | - |
ectoine | mutant S205A, pH 7.5, 32°C | Virgibacillus salexigens | |
7.1 | - |
ectoine | mutant P198A, pH 7.5, 32°C | Virgibacillus salexigens | |
8 | - |
ectoine | mutant F263Y, pH 7.5, 32°C | Virgibacillus salexigens | |
17.3 | - |
ectoine | mutant S165A, pH 7.5, 32°C | Virgibacillus salexigens | |
19.6 | - |
ectoine | mutant F242Y, pH 7.5, 32°C | Virgibacillus salexigens |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Virgibacillus salexigens | Q2TDY4 | - |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ectoine + 2-oxoglutarate + O2 | - |
Virgibacillus salexigens | 5-hydroxyectoine + succinate + CO2 | - |
? |