BRENDA - Enzyme Database
show all sequences of 6.2.1.33

The mechanism of domain alternation in the acyl-adenylate forming ligase superfamily member 4-chlorobenzoate: coenzyme A ligase

Wu, R.; Reger, A.S.; Lu, X.; Gulick, A.M.; Dunaway-Mariano, D.; Biochemistry 48, 4115-4125 (2009)

Data extracted from this reference:

Crystallization (Commentary)
Crystallization (Commentary)
Organism
mutant D402P, to 2.6 A resolution, and comparison with wild-type. The C-terminal domain rotates by about 140 degrees between the two states that catalyze the adenylation and thioester-forming half-reactions. The domain rotation is accompanied by a change in the main chain torsional angles of residue D402; mutant D402P, to 2.6 A resolution, and comparison with wild-type. The C-terminal domain rotates by about 140 degrees between the two states that catalyze the adenylation and thioester-forming half-reactions. The domain rotation is accompanied by a change in the main chain torsional angles of residue D402
Alcaligenes sp. AL3007
Engineering
Protein Variants
Commentary
Organism
D402A
100fold decrease in ratio kcat/Km for CoA
Alcaligenes sp. AL3007
D402P
crystallization data. Mutant adopts the proposed adenylate-forming conformation with very little change to the overall structure.The ability of the mutant to catalyze the adenylate-forming half-reaction is reduced by about 3fold, catalysis of the second half-reaction is reduced by 4 orders of magnitude
Alcaligenes sp. AL3007
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.0009
-
4-Chlorobenzoate
wild-type, pH 7.5, 25°C
Alcaligenes sp. AL3007
0.0012
-
4-Chlorobenzoate
mutant D402P, pH 7.5, 25°C
Alcaligenes sp. AL3007
0.0023
-
4-Chlorobenzoate
mutant D402A, pH 7.5, 25°C
Alcaligenes sp. AL3007
0.0131
-
ATP
mutant D402P, pH 7.5, 25°C
Alcaligenes sp. AL3007
0.0281
-
ATP
mutant D402A, pH 7.5, 25°C
Alcaligenes sp. AL3007
0.124
-
ATP
wild-type, pH 7.5, 25°C
Alcaligenes sp. AL3007
0.31
-
CoA
wild-type, pH 7.5, 25°C
Alcaligenes sp. AL3007
2
-
CoA
mutant D402P, pH 7.5, 25°C
Alcaligenes sp. AL3007
6
-
CoA
mutant D402A, pH 7.5, 25°C
Alcaligenes sp. AL3007
Organism
Organism
UniProt
Commentary
Textmining
Alcaligenes sp. AL3007
-
-
-
Alcaligenes sp. AL3007
Q8GN86
mutant D402P
-
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
Substrate Product ID
4-Chlorobenzoate + CoA + ATP
-
702299
Alcaligenes sp. AL3007
4-Chlorobenzoyl-CoA + AMP + diphosphate
-
-
-
?
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.9
-
4-Chlorobenzoate
mutant D402A, pH 7.5, 25°C
Alcaligenes sp. AL3007
1
-
ATP
mutant D402A, pH 7.5, 25°C
Alcaligenes sp. AL3007
1.8
-
CoA
mutant D402A, pH 7.5, 25°C
Alcaligenes sp. AL3007
2.8
-
ATP
mutant D402P, pH 7.5, 25°C
Alcaligenes sp. AL3007
3.4
-
4-Chlorobenzoate
mutant D402P, pH 7.5, 25°C
Alcaligenes sp. AL3007
3.7
-
CoA
mutant D402P, pH 7.5, 25°C
Alcaligenes sp. AL3007
9.2
-
4-Chlorobenzoate
wild-type, pH 7.5, 25°C
Alcaligenes sp. AL3007
9.3
-
CoA
wild-type, pH 7.5, 25°C
Alcaligenes sp. AL3007
11.5
-
ATP
wild-type, pH 7.5, 25°C
Alcaligenes sp. AL3007
Crystallization (Commentary) (protein specific)
Crystallization
Organism
mutant D402P, to 2.6 A resolution, and comparison with wild-type. The C-terminal domain rotates by about 140 degrees between the two states that catalyze the adenylation and thioester-forming half-reactions. The domain rotation is accompanied by a change in the main chain torsional angles of residue D402
Alcaligenes sp. AL3007
Engineering (protein specific)
Protein Variants
Commentary
Organism
D402A
100fold decrease in ratio kcat/Km for CoA
Alcaligenes sp. AL3007
D402P
crystallization data. Mutant adopts the proposed adenylate-forming conformation with very little change to the overall structure.The ability of the mutant to catalyze the adenylate-forming half-reaction is reduced by about 3fold, catalysis of the second half-reaction is reduced by 4 orders of magnitude
Alcaligenes sp. AL3007
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.0009
-
4-Chlorobenzoate
wild-type, pH 7.5, 25°C
Alcaligenes sp. AL3007
0.0012
-
4-Chlorobenzoate
mutant D402P, pH 7.5, 25°C
Alcaligenes sp. AL3007
0.0023
-
4-Chlorobenzoate
mutant D402A, pH 7.5, 25°C
Alcaligenes sp. AL3007
0.0131
-
ATP
mutant D402P, pH 7.5, 25°C
Alcaligenes sp. AL3007
0.0281
-
ATP
mutant D402A, pH 7.5, 25°C
Alcaligenes sp. AL3007
0.124
-
ATP
wild-type, pH 7.5, 25°C
Alcaligenes sp. AL3007
0.31
-
CoA
wild-type, pH 7.5, 25°C
Alcaligenes sp. AL3007
2
-
CoA
mutant D402P, pH 7.5, 25°C
Alcaligenes sp. AL3007
6
-
CoA
mutant D402A, pH 7.5, 25°C
Alcaligenes sp. AL3007
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ID
4-Chlorobenzoate + CoA + ATP
-
702299
Alcaligenes sp. AL3007
4-Chlorobenzoyl-CoA + AMP + diphosphate
-
-
-
?
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.9
-
4-Chlorobenzoate
mutant D402A, pH 7.5, 25°C
Alcaligenes sp. AL3007
1
-
ATP
mutant D402A, pH 7.5, 25°C
Alcaligenes sp. AL3007
1.8
-
CoA
mutant D402A, pH 7.5, 25°C
Alcaligenes sp. AL3007
2.8
-
ATP
mutant D402P, pH 7.5, 25°C
Alcaligenes sp. AL3007
3.4
-
4-Chlorobenzoate
mutant D402P, pH 7.5, 25°C
Alcaligenes sp. AL3007
3.7
-
CoA
mutant D402P, pH 7.5, 25°C
Alcaligenes sp. AL3007
9.2
-
4-Chlorobenzoate
wild-type, pH 7.5, 25°C
Alcaligenes sp. AL3007
9.3
-
CoA
wild-type, pH 7.5, 25°C
Alcaligenes sp. AL3007
11.5
-
ATP
wild-type, pH 7.5, 25°C
Alcaligenes sp. AL3007
KCat/KM [mM/s]
kcat/KM Value [1/mMs-1]
kcat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
0.02
-
CoA
mutant D402P, pH 7.5, 25°C
Alcaligenes sp. AL3007
0.3
-
CoA
mutant D402A, pH 7.5, 25°C
Alcaligenes sp. AL3007
2
-
ATP
mutant D402P, pH 7.5, 25°C
Alcaligenes sp. AL3007
30
-
4-Chlorobenzoate
mutant D402P, pH 7.5, 25°C
Alcaligenes sp. AL3007
30
-
CoA
wild-type, pH 7.5, 25°C
Alcaligenes sp. AL3007
40
-
ATP
mutant D402A, pH 7.5, 25°C
Alcaligenes sp. AL3007
90
-
ATP
wild-type, pH 7.5, 25°C
Alcaligenes sp. AL3007
400
-
4-Chlorobenzoate
mutant D402A, pH 7.5, 25°C
Alcaligenes sp. AL3007
10000
-
4-Chlorobenzoate
wild-type, pH 7.5, 25°C
Alcaligenes sp. AL3007
KCat/KM [mM/s] (protein specific)
KCat/KM Value [1/mMs-1]
KCat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
0.02
-
CoA
mutant D402P, pH 7.5, 25°C
Alcaligenes sp. AL3007
0.3
-
CoA
mutant D402A, pH 7.5, 25°C
Alcaligenes sp. AL3007
2
-
ATP
mutant D402P, pH 7.5, 25°C
Alcaligenes sp. AL3007
30
-
4-Chlorobenzoate
mutant D402P, pH 7.5, 25°C
Alcaligenes sp. AL3007
30
-
CoA
wild-type, pH 7.5, 25°C
Alcaligenes sp. AL3007
40
-
ATP
mutant D402A, pH 7.5, 25°C
Alcaligenes sp. AL3007
90
-
ATP
wild-type, pH 7.5, 25°C
Alcaligenes sp. AL3007
400
-
4-Chlorobenzoate
mutant D402A, pH 7.5, 25°C
Alcaligenes sp. AL3007
10000
-
4-Chlorobenzoate
wild-type, pH 7.5, 25°C
Alcaligenes sp. AL3007
Other publictions for EC 6.2.1.33
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Synonyms
Temperature Optimum [°C]
Temperature Range [°C]
Temperature Stability [°C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [°C] (protein specific)
Temperature Range [°C] (protein specific)
Temperature Stability [°C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
702299
Wu
The mechanism of domain altern ...
Alcaligenes sp. AL3007
Biochemistry
48
4115-4125
2009
-
-
-
1
2
-
-
9
-
-
-
-
-
2
-
-
-
-
-
-
-
-
2
-
-
-
-
-
9
-
-
-
-
-
-
-
-
-
-
-
2
2
-
-
-
-
9
-
-
-
-
-
-
-
-
-
-
-
-
2
-
-
-
-
9
-
-
-
-
-
-
-
-
9
9
690987
Reger
Structural characterization of ...
Alcaligenes sp.
Biochemistry
47
8016-8025
2008
-
-
-
1
-
-
1
-
-
1
-
1
-
1
-
-
-
1
-
-
-
-
1
1
2
1
-
-
-
1
-
-
1
-
-
-
-
-
-
1
1
-
-
-
1
-
-
-
1
-
1
-
-
-
-
-
-
-
-
1
1
1
-
-
-
1
-
-
-
-
-
-
-
-
-
690988
Wu
Mechanism of 4-chlorobenzoate: ...
Alcaligenes sp.
Biochemistry
47
8026-8039
2008
-
-
1
1
25
-
3
1
-
1
-
1
-
1
-
-
1
1
-
-
-
-
3
1
2
1
-
-
-
1
-
-
1
1
-
-
-
-
1
1
1
25
-
-
3
1
1
-
1
-
1
-
-
-
1
-
-
-
-
3
1
1
-
-
-
1
-
-
-
-
-
-
-
-
-
661151
Gulick
Crystal structure of 4-chlorob ...
Alcaligenes sp.
Biochemistry
43
8670-8679
2004
-
-
-
1
-
-
-
-
-
-
-
1
-
3
-
-
1
-
-
-
-
-
1
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
1
-
-
-
1
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
661288
Zhou
The purification and character ...
Arthrobacter sp., Arthrobacter sp. TM-1
Biodegradation
15
97-109
2004
-
-
-
-
-
-
-
3
-
-
2
-
-
5
-
-
1
-
-
-
1
-
9
1
1
1
1
-
-
1
1
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
3
-
-
2
-
-
-
-
1
-
-
1
-
9
1
1
1
-
-
1
1
-
1
-
-
-
-
-
-
776
Chang
Acyl-adenylate motif of the ac ...
Pseudomonas sp., Pseudomonas sp. CBS3
Biochemistry
36
15650-15659
1997
-
-
-
-
5
-
-
15
-
-
-
-
-
10
-
-
1
1
-
-
-
-
2
-
-
-
-
-
15
-
-
-
-
-
-
-
-
-
-
-
-
5
-
-
-
-
15
-
-
-
-
-
-
-
1
-
-
-
-
2
-
-
-
-
15
-
-
-
-
-
-
-
-
-
-
777
Chang
Determination of the chemical ...
Pseudomonas sp., Pseudomonas sp. CBS3
Biochemistry
35
13478-13484
1996
-
-
-
-
-
-
-
-
-
-
-
2
-
10
-
-
-
1
-
-
-
-
4
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
4
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
781
Dunaway-Mariano
On the origins and functions o ...
Pseudomonas sp.
Biodegradation
5
259-276
1994
-
-
-
-
-
-
-
1
-
1
-
-
-
1
-
-
-
-
-
-
-
-
1
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
1
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
778
Loeffler
Purification and properties of ...
Pseudomonas sp.
Biol. Chem. Hoppe-Seyler
373
1001-1007
1992
-
-
-
-
-
-
4
3
-
6
3
-
-
2
-
-
1
-
-
-
1
2
9
1
-
1
-
1
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
4
-
3
-
6
3
-
-
-
-
1
-
-
1
2
9
1
1
-
1
-
1
-
-
-
-
-
-
-
-
-
779
Loeffler
-
Characterization of 4-chlorobe ...
Pseudomonas sp.
DECHEMA Biotechnol. Conf. (Pt. A. Microbial Principles in Bioprocesses: Cell Culture Technology - Downstream Processing and Recovery)
5
25-28
1992
-
-
-
-
-
-
6
3
-
-
-
1
-
1
-
-
-
-
-
-
-
-
9
-
-
1
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
6
-
3
-
-
-
1
-
-
-
-
-
-
-
-
9
-
1
-
-
-
1
-
-
-
-
-
-
-
-
-
780
Chang
Isolation and characterization ...
Pseudomonas sp.
Biochemistry
31
5605-5610
1992
-
-
1
-
-
-
-
3
-
3
2
-
-
3
-
-
1
-
-
-
-
-
6
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
3
-
3
2
-
-
-
-
1
-
-
-
-
6
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-