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Literature summary for 2.1.1.164 extracted from

  • Singh, S.; McCoy, J.G.; Zhang, C.; Bingman, C.A.; Phillips, G.N. Jr.; Thorson, J.S.
    Structure and mechanism of the rebeccamycin sugar 4'-O-methyltransferase RebM (2008), J. Biol. Chem., 283, 22628-22636.
    View publication on PubMedView publication on EuropePMC

Crystallization (Commentary)

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

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 [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/ Products (Substrates)

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

Organism UniProt Comment Textmining
Lentzea aerocolonigenes Q8KZ94
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4'-demethylrebeccamycin + S-adenosyl-L-methionine
-
Lentzea aerocolonigenes rebeccamycin + S-adenosyl-L-homocysteine
-
?

Synonyms

Synonyms Comment Organism
RebM
-
Lentzea aerocolonigenes

Turnover Number [1/s]

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 [mM/s]

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