Crystallization (Comment) | Organism |
---|---|
hanging drop vapor diffusion method at 20°C. The 2.65 A crystal structure of the rebeccamycin 4'-O-methyltransferase RebM in complex with S-adenosyl-L-homocysteine reveals RebM to adopt a typical S-adenosyl methionine binding fold of small molecule O-methyltransferases and display a weak dimerization domain unique to methyltransferases | Lentzea aerocolonigenes |
Protein Variants | Comment | Organism |
---|---|---|
C70A | kcat for S-adenosyl-L-methionine is nearly identical to wild-type value | Lentzea aerocolonigenes |
C70S | kcat for S-adenosyl-L-methionine is 1.53fold higher than wild-type value | Lentzea aerocolonigenes |
D166A | kcat for S-adenosyl-L-methionine is 10% of wild-type value | Lentzea aerocolonigenes |
H140A | kcat for S-adenosyl-L-methionine is 5% of wild-type value | Lentzea aerocolonigenes |
H141A | kcat for S-adenosyl-L-methionine is 21% of wild-type value | Lentzea aerocolonigenes |
H149A/H141A | activity is below the detection limit | Lentzea aerocolonigenes |
L136V | kcat for S-adenosyl-L-methionine is 54% of wild-type value | Lentzea aerocolonigenes |
P75S | although properly folded based upon CD spectroscopy, the mutant displays a substantially reduced affinity for AdoMet (about 10fold increase in Km). kcat for S-adenosyl-L-methionine is 13% of wild-type value | Lentzea aerocolonigenes |
S138A | kcat for S-adenosyl-L-methionine is 49% of wild-type value | Lentzea aerocolonigenes |
W134Y | kcat for S-adenosyl-L-methionine is 35% of wild-type value | Lentzea aerocolonigenes |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0112 | - |
4'-demethylrebeccamycin | pH 8.0, 25°C, mutant enzyme H140A | Lentzea aerocolonigenes | |
0.012 | - |
S-adenosyl-L-methionine | pH 8.0, 25°C, wild-type enzyme | Lentzea aerocolonigenes | |
0.016 | - |
4'-demethylrebeccamycin | pH 8.0, 25°C, mutant enzyme W134Y | Lentzea aerocolonigenes | |
0.017 | - |
S-adenosyl-L-methionine | pH 8.0, 25°C, mutant enzyme C70A | Lentzea aerocolonigenes | |
0.017 | - |
4'-demethylrebeccamycin | pH 8.0, 25°C, mutant enzyme C70S | Lentzea aerocolonigenes | |
0.018 | - |
S-adenosyl-L-methionine | pH 8.0, 25°C, mutant enzyme S138A | Lentzea aerocolonigenes | |
0.021 | - |
4'-demethylrebeccamycin | pH 8.0, 25°C, mutant enzyme S138A | Lentzea aerocolonigenes | |
0.0215 | - |
4'-demethylrebeccamycin | pH 8.0, 25°C, mutant enzyme L136V | Lentzea aerocolonigenes | |
0.0225 | - |
4'-demethylrebeccamycin | pH 8.0, 25°C, mutant enzyme C70A | Lentzea aerocolonigenes | |
0.0311 | - |
S-adenosyl-L-methionine | pH 8.0, 25°C, mutant enzyme H140A | Lentzea aerocolonigenes | |
0.032 | - |
4'-demethylrebeccamycin | pH 8.0, 25°C, mutant enzyme D166A | Lentzea aerocolonigenes | |
0.032 | - |
S-adenosyl-L-methionine | pH 8.0, 25°C, mutant enzyme W134Y | Lentzea aerocolonigenes | |
0.0345 | - |
S-adenosyl-L-methionine | pH 8.0, 25°C, mutant enzyme C70S | Lentzea aerocolonigenes | |
0.0355 | - |
4'-demethylrebeccamycin | pH 8.0, 25°C, wild-type enzyme | Lentzea aerocolonigenes | |
0.0535 | - |
4'-demethylrebeccamycin | pH 8.0, 25°C, mutant enzyme P75S | Lentzea aerocolonigenes | |
0.056 | - |
S-adenosyl-L-methionine | pH 8.0, 25°C, mutant enzyme L136V | Lentzea aerocolonigenes | |
0.0798 | - |
4'-demethylrebeccamycin | pH 8.0, 25°C, mutant enzyme H141A | Lentzea aerocolonigenes | |
0.0943 | - |
S-adenosyl-L-methionine | pH 8.0, 25°C, mutant enzyme H141A | Lentzea aerocolonigenes | |
0.162 | - |
S-adenosyl-L-methionine | pH 8.0, 25°C, mutant enzyme D166A | Lentzea aerocolonigenes | |
0.162 | - |
S-adenosyl-L-methionine | pH 8.0, 25°C, mutant enzyme P75S | Lentzea aerocolonigenes |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
4'-demethylrebeccamycin + S-adenosyl-L-methionine | Lentzea aerocolonigenes | - |
rebeccamycin + S-adenosyl-L-homocysteine | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Lentzea aerocolonigenes | Q8KZ94 | - |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
4'-demethylrebeccamycin + S-adenosyl-L-methionine | - |
Lentzea aerocolonigenes | rebeccamycin + S-adenosyl-L-homocysteine | - |
? |
Synonyms | Comment | Organism |
---|---|---|
RebM | - |
Lentzea aerocolonigenes |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0023 | - |
S-adenosyl-L-methionine | pH 8.0, 25°C, mutant enzyme H140A | Lentzea aerocolonigenes | |
0.005 | - |
S-adenosyl-L-methionine | pH 8.0, 25°C, mutant enzyme D166A | Lentzea aerocolonigenes | |
0.0063 | - |
S-adenosyl-L-methionine | pH 8.0, 25°C, mutant enzyme P75S | Lentzea aerocolonigenes | |
0.01 | - |
S-adenosyl-L-methionine | pH 8.0, 25°C, mutant enzyme H141A | Lentzea aerocolonigenes | |
0.017 | - |
S-adenosyl-L-methionine | pH 8.0, 25°C, mutant enzyme W134Y | Lentzea aerocolonigenes | |
0.023 | - |
S-adenosyl-L-methionine | pH 8.0, 25°C, mutant enzyme S138A | Lentzea aerocolonigenes | |
0.025 | - |
S-adenosyl-L-methionine | pH 8.0, 25°C, mutant enzyme L136V | Lentzea aerocolonigenes | |
0.047 | - |
S-adenosyl-L-methionine | pH 8.0, 25°C, wild-type enzyme | Lentzea aerocolonigenes | |
0.048 | - |
S-adenosyl-L-methionine | pH 8.0, 25°C, mutant enzyme C70A | Lentzea aerocolonigenes | |
0.072 | - |
S-adenosyl-L-methionine | pH 8.0, 25°C, mutant enzyme C70S | Lentzea aerocolonigenes |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.031 | - |
S-adenosyl-L-methionine | pH 8.0, 25°C, mutant enzyme D166A | Lentzea aerocolonigenes | |
0.031 | - |
S-adenosyl-L-methionine | pH 8.0, 25°C, mutant enzyme H140A/H141A | Lentzea aerocolonigenes | |
0.039 | - |
S-adenosyl-L-methionine | pH 8.0, 25°C, mutant enzyme P75S | Lentzea aerocolonigenes | |
0.074 | - |
S-adenosyl-L-methionine | pH 8.0, 25°C, mutant enzyme H140A | Lentzea aerocolonigenes | |
0.11 | - |
S-adenosyl-L-methionine | pH 8.0, 25°C, mutant enzyme H141A | Lentzea aerocolonigenes | |
0.45 | - |
S-adenosyl-L-methionine | pH 8.0, 25°C, mutant enzyme L136V | Lentzea aerocolonigenes | |
0.53 | - |
S-adenosyl-L-methionine | pH 8.0, 25°C, mutant enzyme W134Y | Lentzea aerocolonigenes | |
1.28 | - |
S-adenosyl-L-methionine | pH 8.0, 25°C, mutant enzyme S138A | Lentzea aerocolonigenes | |
2.09 | - |
S-adenosyl-L-methionine | pH 8.0, 25°C, mutant enzyme C70S | Lentzea aerocolonigenes | |
2.8 | - |
S-adenosyl-L-methionine | pH 8.0, 25°C, mutant enzyme C70A | Lentzea aerocolonigenes | |
3.9 | - |
S-adenosyl-L-methionine | pH 8.0, 25°C, wild-type enzyme | Lentzea aerocolonigenes |