Crystallization (Comment) | Organism |
---|---|
molecular dynamics simulations of wild-type SsuD and variant enzymes bound with different combinations of FMNH2, C4a-peroxyflavin intermediate FMNOO-, and octanesulfonate. Mobile loop conformations are open, closed, and semiclosed. The substrate-free SsuD system has a wide opening capable of providing full access for substrates to enter the active site. Upon binding FMNH2, SsuD adopts a closed conformation. Salt bridges, Asp111-Arg263 and Glu205-Arg271, are particularly important in maintaining the closed conformation. With both FMNH2 and octanesulfonate bound in SsuD, a second conformation is formed dependent upon a favorable pi-pi interaction between residues His124 and Phe261 | Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
F261W | complete loss of activity | Escherichia coli |
R263A | similar kinetic parameters as wild-type | Escherichia coli |
R263A/R271A | similar kinetic parameters as wild-type | Escherichia coli |
R271A | mutation does not alter the catalytic activity, but in the presence of reduced flavin the variant is more susceptible to proteolytic digestion compared to wild-type | Escherichia coli |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.05 | - |
octanesulfonate | wild-type, pH 7.5, 25°C | Escherichia coli | |
0.057 | - |
octanesulfonate | mutant R271A, pH 7.5, 25°C | Escherichia coli | |
0.113 | - |
octanesulfonate | mutant R263A/R271A, pH 7.5, 25°C | Escherichia coli | |
0.235 | - |
octanesulfonate | mutant R263A, pH 7.5, 25°C | Escherichia coli |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | P80645 | - |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
octanesulfonate + FMNH2 + O2 | - |
Escherichia coli | octanal + FMN + sulfite + H2O | - |
? |
Synonyms | Comment | Organism |
---|---|---|
SsuD | - |
Escherichia coli |
YcbN | - |
Escherichia coli |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.84 | - |
octanesulfonate | wild-type, pH 7.5, 25°C | Escherichia coli | |
1.03 | - |
octanesulfonate | mutant R263A, pH 7.5, 25°C | Escherichia coli | |
1.1 | - |
octanesulfonate | mutant R263A/R271A, pH 7.5, 25°C | Escherichia coli | |
1.4 | - |
octanesulfonate | mutant R271A, pH 7.5, 25°C | Escherichia coli |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
4.4 | - |
octanesulfonate | mutant R263A, pH 7.5, 25°C | Escherichia coli | |
10 | - |
octanesulfonate | mutant R263A/R271A, pH 7.5, 25°C | Escherichia coli | |
17 | - |
octanesulfonate | wild-type, pH 7.5, 25°C | Escherichia coli | |
25 | - |
octanesulfonate | mutant R271A, pH 7.5, 25°C | Escherichia coli |