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Literature summary for 1.5.1.3 extracted from

  • Boeck, R.A.; Soulages, J.L.; Barrow, W.W.
    Substrate and inhibitor specificity of Mycobacterium avium dihydrofolate reductase (2007), FEBS J., 274, 3286-3298.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli Mycobacterium avium

Protein Variants

Protein Variants Comment Organism
D31A 7% of wild-type activity Mycobacterium avium
D31E 18.9% of wild-type activity Mycobacterium avium
D31L 0.39% of wild-type activity Mycobacterium avium
D31N 15.4% of wild-type activity Mycobacterium avium
D31Q 14.7% of wild-type activity Mycobacterium avium
L32A 96% of wild-type activity. 12-fold decrease in affinity for trimethoprim Mycobacterium avium
L32D 33% of wild-type activity. 7-fold decrease in affinity for trimethoprim Mycobacterium avium
L32F 103% of wild-type activity Mycobacterium avium
V76A 98.7% of wild-type activity Mycobacterium avium

Inhibitors

Inhibitors Comment Organism Structure
trimethoprim
-
Mycobacterium avium

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.00065
-
NADPH pH 7.0, 30°C, mutant D31Q Mycobacterium avium
0.00068
-
NADPH pH 7.0, 30°C, mutant D31A Mycobacterium avium
0.00068
-
7,8-dihydrofolate pH 7.0, 30°C, mutant L32F Mycobacterium avium
0.0007
-
7,8-dihydrofolate pH 7.0, 30°C, wild-type Mycobacterium avium
0.00077
-
NADPH pH 7.0, 30°C, mutant D31N Mycobacterium avium
0.00078
-
7,8-dihydrofolate pH 7.0, 30°C, mutant V76A Mycobacterium avium
0.00084
-
NADPH pH 7.0, 30°C, mutant L32D Mycobacterium avium
0.00096
-
7,8-dihydrofolate pH 7.0, 30°C, mutant L32A Mycobacterium avium
0.0013
-
NADPH pH 7.0, 30°C, mutant L32F Mycobacterium avium
0.0014
-
NADPH pH 7.0, 30°C, mutant D31L Mycobacterium avium
0.0014
-
NADPH pH 7.0, 30°C, mutant L32A Mycobacterium avium
0.0015
-
NADPH pH 7.0, 30°C, mutant V76A Mycobacterium avium
0.0015
-
NADPH pH 7.0, 30°C, wild-type Mycobacterium avium
0.0019
-
7,8-dihydrofolate pH 7.0, 30°C, mutant D31E Mycobacterium avium
0.002
-
NADPH pH 7.0, 30°C, mutant D31E Mycobacterium avium
0.0021
-
7,8-dihydrofolate pH 7.0, 30°C, mutant D31N Mycobacterium avium
0.0023
-
7,8-dihydrofolate pH 7.0, 30°C, mutant D31Q Mycobacterium avium
0.0051
-
7,8-dihydrofolate pH 7.0, 30°C, mutant L32D Mycobacterium avium
0.037
-
7,8-dihydrofolate pH 7.0, 30°C, mutant D31A Mycobacterium avium
0.198
-
7,8-dihydrofolate pH 7.0, 30°C, mutant D31L Mycobacterium avium

Organism

Organism UniProt Comment Textmining
Mycobacterium avium O30463
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
7,8-dihydrofolate + NADPH + H+
-
Mycobacterium avium 5,6,7,8-tetrahydrofolate + NADP+
-
?

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.02
-
NADPH pH 7.0, 30°C, mutant D31L Mycobacterium avium
0.025
-
7,8-dihydrofolate pH 7.0, 30°C, mutant D31L Mycobacterium avium
0.59
-
NADPH pH 7.0, 30°C, mutant D31A Mycobacterium avium
0.86
-
7,8-dihydrofolate pH 7.0, 30°C, mutant D31A Mycobacterium avium
1.5
-
7,8-dihydrofolate pH 7.0, 30°C, mutant D31Q Mycobacterium avium
2 3.7 NADPH pH 7.0, 30°C, mutant V76A Mycobacterium avium
2.05
-
NADPH pH 7.0, 30°C, mutant D31E Mycobacterium avium
2.18
-
NADPH pH 7.0, 30°C, mutant D31Q Mycobacterium avium
3.58
-
7,8-dihydrofolate pH 7.0, 30°C, mutant D31E Mycobacterium avium
5.03
-
7,8-dihydrofolate pH 7.0, 30°C, mutant D31N Mycobacterium avium
7.2
-
7,8-dihydrofolate pH 7.0, 30°C, mutant L32D Mycobacterium avium
8.5
-
NADPH pH 7.0, 30°C, mutant D31N Mycobacterium avium
10
-
NADPH pH 7.0, 30°C, mutant L32D Mycobacterium avium
16.2
-
7,8-dihydrofolate pH 7.0, 30°C, mutant L32A Mycobacterium avium
17.8
-
NADPH pH 7.0, 30°C, mutant L32A Mycobacterium avium
18.7
-
7,8-dihydrofolate pH 7.0, 30°C, mutant V76A Mycobacterium avium
20.5
-
7,8-dihydrofolate pH 7.0, 30°C, wild-type Mycobacterium avium
21
-
7,8-dihydrofolate pH 7.0, 30°C, mutant L32F Mycobacterium avium
23.2
-
NADPH pH 7.0, 30°C, mutant L32F Mycobacterium avium
26.8
-
NADPH pH 7.0, 30°C, wild-type Mycobacterium avium