Cloned (Comment) | Organism |
---|---|
recombinant expression of His-tagged wild-type and selenomethionine-labeled enzymes in Escherichia coli strain BL21(DE3) and B834(DE3), respectively | Saccharomyces cerevisiae |
Crystallization (Comment) | Organism |
---|---|
purified recombinant selenomethionine-labeled enzyme and of purified recombinant wild-type enzyme in complex with cofactor pyridoxal 5'-phosphate, hanging drop vapor diffusion method, mixing of 0.001 ml of 10 mg/ml protein in 100 mM NaCl, 20 mM Tris-Cl, pH 8.0, with an equal volume of reservoir solution containing 20% w/v PEG monomethyl ether 5000, 0.1 M Bis-Tris, pH 6.2, 1-2 days to 1 week, 16°C, X-ray diffraction structure determination and analysis at 1.90-2.20 A resolution | Saccharomyces cerevisiae |
Protein Variants | Comment | Organism |
---|---|---|
K180A | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Saccharomyces cerevisiae |
N360A | site-directed mutagenesis, inactive mutant | Saccharomyces cerevisiae |
N360D | site-directed mutagenesis, inactive mutant | Saccharomyces cerevisiae |
T30A | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Saccharomyces cerevisiae |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
4-amino-4-deoxychorismate | Saccharomyces cerevisiae | - |
4-aminobenzoate + pyruvate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Saccharomyces cerevisiae | Q03266 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged wild-type and selenomethionine-labeled enzymes from Escherichia coli strain BL21(DE3) and B834(DE3), respectively, by nickel affinity chromatography | Saccharomyces cerevisiae |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
4-amino-4-deoxychorismate = 4-aminobenzoate + pyruvate | catalytic mechanism, overview | Saccharomyces cerevisiae |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
4-amino-4-deoxychorismate | - |
Saccharomyces cerevisiae | 4-aminobenzoate + pyruvate | - |
? | |
4-amino-4-deoxychorismate | simulation and validation of the substrate-binding model, overview | Saccharomyces cerevisiae | 4-aminobenzoate + pyruvate | - |
? |
Subunits | Comment | Organism |
---|---|---|
monomer | - |
Saccharomyces cerevisiae |
Synonyms | Comment | Organism |
---|---|---|
4-amino-4-deoxychorismate lyase | - |
Saccharomyces cerevisiae |
Abz2 | - |
Saccharomyces cerevisiae |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
pyridoxal 5'-phosphate | dependent on, one molecule of cofactor is deeply buried in the cleft between domains I and II, pyridoxal 5'-phosphate adopts the re-face specificity facing the protein side and is covalently linked to the catalytic residue Lys251 by forming an internal aldimine bond, Schiff base linkage | Saccharomyces cerevisiae |
General Information | Comment | Organism |
---|---|---|
evolution | 4-amino-4-deoxychorismate lyases can be divided into two classes of dimeric and monomeric enzyme, respectively | Saccharomyces cerevisiae |
additional information | the catalytic residue Lys251 covalently binds the cofactor pyridoxal 5'-phosphate | Saccharomyces cerevisiae |