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Literature summary for 1.1.1.262 extracted from

  • Sivaraman, J.; Li, Y.; Banks, J.; Cane, D.E.; Matte, A.; Cygler, M.
    Crystal structure of Escherichia coli PdxA, an enzyme involved in the pyridoxal phosphate biosynthesis pathway (2003), J. Biol. Chem., 278, 43682-43690.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
gene pdxA, expression of the His-tagged enzyme with a thrombin cleavage site in Escherichia coli Escherichia coli

Crystallization (Commentary)

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

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/ Products (Substrates)

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

Organism UniProt Comment Textmining
Escherichia coli P19624
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged enzyme from Escherichia coli by nickel affinity chromatography, cleavage by thrombin Escherichia coli

Reaction

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 and Products (Substrate)

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

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

Synonyms Comment Organism
PdxA
-
Escherichia coli

Cofactor

Cofactor Comment Organism Structure
NAD(P)+ putative binding site structure Escherichia coli