Cloned (Comment) | Organism |
---|---|
gene pdxA, expression of the His-tagged enzyme with a thrombin cleavage site in Escherichia coli | Escherichia coli |
Crystallization (Comment) | Organism |
---|---|
8.1 mg/ml purified recombinant enzyme in complex with substrate 4-(phosphonooxy)threonine and Zn2+ in 20 mM Tris-HCl, pH 8.0, 0.2 M NaCl, 10 mM DTT, 5% w/v glycerol, hanging drop vapour diffusion method, 0.002 ml protein solution with 0.004 ml reservoir solution, usage of 2 reservoir solution variants resulting in 2 differen crystal forms, solution 1 contains 7.5% w/v PEG 8000, 0.1 M NaOAc, pH 5.5, 10 mM MgCl2, 10 mM NaKHPO4, pH 5.9, solution 2 contains 20% w/v PEG 8000, 100 mM HEPES, pH 7.5, 75 mM citrate, and 100 mM MgCl2, suspension over reservoir solution, X-ray diffraction structure determination and analysis at 2,0 and 2.45 A resolution, respectively | Escherichia coli |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Zn2+ | divalent metal ion-dependent enzyme, Zn2+ is bound to the active site of each monomer coordinated by 3 histidine residues from both monomer | Escherichia coli |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
4-(phosphonooxy)threonine + NAD(P)+ | Escherichia coli | fourth step of the pyridoxal 5'-phosphate biosynthesis | 2-amino-3-oxo-4-phosphonooxybutyrate + NAD(P)H | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | P19624 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged enzyme from Escherichia coli by nickel affinity chromatography, cleavage by thrombin | Escherichia coli |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
4-phosphooxy-L-threonine + NAD+ = 3-amino-2-oxopropyl phosphate + CO2 + NADH + H+ | Asp247 and Asp267 are involved in formation and maintaining of the integrity of the active site | Escherichia coli |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2-amino-3-oxo-4-phosphonooxybutyrate + H2O | step 2 of the overall reaction | Escherichia coli | 3-amino-1-hydroxyacetone 1-phosphate + CO2 | i.e. AHAP | ? | |
4-(phosphonooxy)threonine + NAD(P)+ | fourth step of the pyridoxal 5'-phosphate biosynthesis | Escherichia coli | 2-amino-3-oxo-4-phosphonooxybutyrate + NAD(P)H | - |
? | |
4-(phosphonooxy)threonine + NAD(P)+ | i.e. 4-hydroxy-L-threonine phosphate or HTP, enzyme strictly requires the phosphate ester form of the substrate, interaction between enzyme and substrate via phosphate group, step 1 of the overall reaction | Escherichia coli | 2-amino-3-oxo-4-phosphonooxybutyrate + NAD(P)H | reaction intermediate | ? |
Subunits | Comment | Organism |
---|---|---|
dimer | tightly bound, monomer fold is alpha/beta/alpha divided into 2 subdomains, the active site is located at the dimer interface | Escherichia coli |
Synonyms | Comment | Organism |
---|---|---|
PdxA | - |
Escherichia coli |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD(P)+ | putative binding site structure | Escherichia coli |