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A structural basis for the mechanism of aspartate-beta-semialdehyde dehydrogenase from Vibrio cholerae

Blanco, J.; Moore, R.A.; Kabaleeswaran, V.; Viola, R.E.; Protein Sci. 12, 27-33 (2003)

Data extracted from this reference:

Application
EC Number
Application
Commentary
Organism
1.2.1.11
additional information
enzyme is an attractive target for development of antibacterial, fungicidal, or herbicidal compounds
Vibrio cholerae
Crystallization (Commentary)
EC Number
Crystallization
Organism
1.2.1.11
12 mg/ml purified recombinant enzyme free or in ternary complex with NADP+ and covalently bound inhibitor S-methyl-L-cysteine sulfoxide, protein in 10 mM HEPES, pH 7.0, 1 mM EDTA, and 1 mM DTT, hanging drop vapour diffusion method, 20°C, with or without 5 mM NADP+ and 5 mM inhibitor, against an equal volume of precipitant solution: containing 18% PEG 3350, 0.2 M sodium acetate, and 0.1 M Tris, pH 8.5 for the free enzyme, or containing 22% PEG 3350, 0.2 M sodium acetate, and 0.1 M sodium citrate, pH 5.6 for the ternary complex, addition of 20% glycerol for crystal freezing, X-ray diffraction structure determination and analysis at 2.8 A and 1.84 A resolution, respectively
Vibrio cholerae
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
1.2.1.11
L-cystine
inactivation, reversible by addition of DTT or 2-mercaptoethanol
Vibrio cholerae
1.2.1.11
S-methyl-L-cysteine sulfoxide
covalently binding inhibitor via Cys134 at the active site, inactivation, inhibition and binding mechanism, reversible by addition of DTT or 2-mercaptoethanol
Vibrio cholerae
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
1.2.1.11
L-4-aspartyl phosphate + NADPH
Vibrio cholerae
reductive dephosphorylation in the aspartate biosynthetic pathway of plants and microorganisms
L-aspartate-4-semialdehyde + phosphate + NADP+
-
-
r
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
1.2.1.11
Vibrio cholerae
Q9KQG2
El Tor N16961, TIGR locus VC2036
-
Reaction
EC Number
Reaction
Commentary
Organism
1.2.1.11
L-aspartate 4-semialdehyde + phosphate + NADP+ = L-4-aspartyl phosphate + NADPH + H+
catalytic mechanism, active site structure, catalytic nucleophile is Cys134
Vibrio cholerae
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.2.1.11
L-4-aspartyl phosphate + NADPH
reductive dephosphorylation in the aspartate biosynthetic pathway of plants and microorganisms
657298
Vibrio cholerae
L-aspartate-4-semialdehyde + phosphate + NADP+
-
-
-
r
1.2.1.11
L-aspartate-4-semialdehyde + phosphate + NADP+
-
657298
Vibrio cholerae
L-4-aspartyl phosphate + NADPH
reverse reaction: reductive dephosphorylation
-
-
r
Subunits
EC Number
Subunits
Commentary
Organism
1.2.1.11
dimer
crystal structure, communication mechanism between the active sites of the subunits, overview
Vibrio cholerae
Cofactor
EC Number
Cofactor
Commentary
Organism
Structure
1.2.1.11
NADP+
binding mechanism and structure
Vibrio cholerae
1.2.1.11
NADPH
-
Vibrio cholerae
Application (protein specific)
EC Number
Application
Commentary
Organism
1.2.1.11
additional information
enzyme is an attractive target for development of antibacterial, fungicidal, or herbicidal compounds
Vibrio cholerae
Cofactor (protein specific)
EC Number
Cofactor
Commentary
Organism
Structure
1.2.1.11
NADP+
binding mechanism and structure
Vibrio cholerae
1.2.1.11
NADPH
-
Vibrio cholerae
Crystallization (Commentary) (protein specific)
EC Number
Crystallization
Organism
1.2.1.11
12 mg/ml purified recombinant enzyme free or in ternary complex with NADP+ and covalently bound inhibitor S-methyl-L-cysteine sulfoxide, protein in 10 mM HEPES, pH 7.0, 1 mM EDTA, and 1 mM DTT, hanging drop vapour diffusion method, 20°C, with or without 5 mM NADP+ and 5 mM inhibitor, against an equal volume of precipitant solution: containing 18% PEG 3350, 0.2 M sodium acetate, and 0.1 M Tris, pH 8.5 for the free enzyme, or containing 22% PEG 3350, 0.2 M sodium acetate, and 0.1 M sodium citrate, pH 5.6 for the ternary complex, addition of 20% glycerol for crystal freezing, X-ray diffraction structure determination and analysis at 2.8 A and 1.84 A resolution, respectively
Vibrio cholerae
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
1.2.1.11
L-cystine
inactivation, reversible by addition of DTT or 2-mercaptoethanol
Vibrio cholerae
1.2.1.11
S-methyl-L-cysteine sulfoxide
covalently binding inhibitor via Cys134 at the active site, inactivation, inhibition and binding mechanism, reversible by addition of DTT or 2-mercaptoethanol
Vibrio cholerae
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
1.2.1.11
L-4-aspartyl phosphate + NADPH
Vibrio cholerae
reductive dephosphorylation in the aspartate biosynthetic pathway of plants and microorganisms
L-aspartate-4-semialdehyde + phosphate + NADP+
-
-
r
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.2.1.11
L-4-aspartyl phosphate + NADPH
reductive dephosphorylation in the aspartate biosynthetic pathway of plants and microorganisms
657298
Vibrio cholerae
L-aspartate-4-semialdehyde + phosphate + NADP+
-
-
-
r
1.2.1.11
L-aspartate-4-semialdehyde + phosphate + NADP+
-
657298
Vibrio cholerae
L-4-aspartyl phosphate + NADPH
reverse reaction: reductive dephosphorylation
-
-
r
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
1.2.1.11
dimer
crystal structure, communication mechanism between the active sites of the subunits, overview
Vibrio cholerae