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Literature summary extracted from

  • Puehringer, S.; RoseFigura, J.; Metlitzky, M.; Toyama, H.; Klinman, J.P.; Schwarzenbacher, R.
    Structural studies of mutant forms of the PQQ-forming enzyme PqqC in the presence of product and substrate (2010), Proteins, 78, 2554-2562.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.3.3.11 gene pqqC, expression of wild-type and mutant enzymes as His-tagged enzyme in Escherichia coli strain BL21(DE3) Klebsiella pneumoniae

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
1.3.3.11 purified recombinant enzyme mutants H154S/PQQ Y175F/PQQ Y175S/R179S in complex with intermediate 3a-(2-amino-2-carboxyethyl)-4,5-dioxo-4,5,6,7,8,9-hexahydroquinoline-7,9-dicarboxylic acid, sitting drop-vapor diffusion method, optimized conditions for each mutant: 0.1 M HEPES, pH 7.0, 0.5% w/v PEG 8000 for mutant H154S/PQQ complex, 0.2 M ammonium sulfate, 0.1 M Bis-Tris, pH 6.5, 25% w/v PEGl 3350 for mutant Y175F/PQQ complex, and 0.2 M sodium chloride, 0.1 M Tris, pH 8.5, 25% w/v PEG 3350 for mutant R179S/Y175S/intermediate complex, all at 20°C, X-ray diffraction structure determination and analysis at 1.3-2.35 A resolution Klebsiella pneumoniae

Protein Variants

EC Number Protein Variants Comment Organism
1.3.3.11 H154S site-directed mutagenesis, active site mutant, even with substrate PQQ bound, the enzyme is still in the open conformation with helices alpha5b and alpha6 unfolded and the active site solvent accessible, no product formation Klebsiella pneumoniae
1.3.3.11 R179S/Y175S site-directed mutagenesis, active site mutant, the mutant shows an open conformation with a reaction intermediate trapped in the active site, the intermediate is tricyclic but nonplanar, implying that it has not undergone oxidatio, R179S/Y175S shows acceptable substrate complex crystals Klebsiella pneumoniae
1.3.3.11 Y175F site-directed mutagenesis, active site mutant, the mutant shows a closed conformation indicating that Y175 is not required for the conformational change, no product formation Klebsiella pneumoniae

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.3.3.11 additional information PqqC is a metal free oxidase Klebsiella pneumoniae

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.3.3.11 28910
-
 x * 28910 Klebsiella pneumoniae

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.3.3.11 6-(2-amino-2-carboxyethyl)-7,8-dioxo-1,2,3,4,7,8-hexahydroquinoline-2,4-dicarboxylate + 3 O2 Klebsiella pneumoniae
-
 4,5-dioxo-1H-pyrrolo[2,3-f]quinoline-2,7,9-tricarboxylate + 2 H2O2 + 2 H2O
-
 ?
1.3.3.11 6-(2-amino-2-carboxyethyl)-7,8-dioxo-1,2,3,4,7,8-hexahydroquinoline-2,4-dicarboxylate + 3 O2 Klebsiella pneumoniae MGH 78578
-
 4,5-dioxo-1H-pyrrolo[2,3-f]quinoline-2,7,9-tricarboxylate + 2 H2O2 + 2 H2O
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.1.5.2 Klebsiella pneumoniae
-
-
-
1.1.5.2 Klebsiella pneumoniae MGH 78578
-
-
-
1.3.3.11 Klebsiella pneumoniae A6T9H1 subsp. pneumoniae
-
1.3.3.11 Klebsiella pneumoniae MGH 78578 A6T9H1 subsp. pneumoniae
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.3.3.11 recombinant His-tagged wild-type and mutant enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration Klebsiella pneumoniae

Reaction

EC Number Reaction Comment Organism Reaction ID
1.3.3.11 6-(2-amino-2-carboxyethyl)-7,8-dioxo-1,2,3,4,7,8-hexahydroquinoline-2,4-dicarboxylate + 3 O2 = 4,5-dioxo-4,5-dihydro-1H-pyrrolo[2,3-f]quinoline-2,7,9-tricarboxylate + 2 H2O2 + 2 H2O reaction mechanism consisting of a series of base catalyzed proton abstractions followed by quinoid/quinol tautomerizations and oxidations, overview Klebsiella pneumoniae
a

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.3.3.11 6-(2-amino-2-carboxyethyl)-7,8-dioxo-1,2,3,4,7,8-hexahydroquinoline-2,4-dicarboxylate + 3 O2
-
 Klebsiella pneumoniae 4,5-dioxo-1H-pyrrolo[2,3-f]quinoline-2,7,9-tricarboxylate + 2 H2O2 + 2 H2O
-
 ?
1.3.3.11 6-(2-amino-2-carboxyethyl)-7,8-dioxo-1,2,3,4,7,8-hexahydroquinoline-2,4-dicarboxylate + 3 O2
-
 Klebsiella pneumoniae MGH 78578 4,5-dioxo-1H-pyrrolo[2,3-f]quinoline-2,7,9-tricarboxylate + 2 H2O2 + 2 H2O
-
 ?

Subunits

EC Number Subunits Comment Organism
1.3.3.11 ? x * 28910 Klebsiella pneumoniae
1.3.3.11 More the enzyme folds into a compact seven-helix bundle, which provide the scaffold for an active site cavity Klebsiella pneumoniae

Synonyms

EC Number Synonyms Comment Organism
1.1.5.2 PqqC
-
 Klebsiella pneumoniae
1.3.3.11 PqqC
-
 Klebsiella pneumoniae
1.3.3.11 pyrroloquinoline quinone synthase C
-
 Klebsiella pneumoniae

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.5.2 pyrroloquinoline quinone i.e. 4,5-dihydro-4,5-dioxo-1H-pyrrolo[2,3-f]quinoline-2,7,9-tricarboxylic acid, prosthetic group, not covalently linked to the polypeptide chain or posttranslationally derived from precursor amino acid residues in the active site of the constituent. enzyme. Biosynthesis of the cofactor in Klebsiella pneumoniae is facilitated by six genes, pqqABCDEF. PqqC is one of two metal free oxidases of known structure and catalyzes the last step of PQQ biogenesis which involves a ring closure and an eight-electron oxidation of the substrate 3a-(2-amino-2-carboxyethyl)-4,5-dioxo-4,5,6,7,8,9-hexahydroquinoline-7,9-dicarboxylic acid, overview Klebsiella pneumoniae

General Information

EC Number General Information Comment Organism
1.3.3.11 metabolism PqqC catalyzes the last step of pyrroloquinoline quinone biogenesis which involves a ring closure and an eight-electron oxidation of the substrate 3a-(2-amino-2-carboxyethyl)-4,5-dioxo-4,5,6,7,8,9-hexahydroquinoline-7,9-dicarboxylic acid Klebsiella pneumoniae
1.3.3.11 additional information the enzyme has 14 conserved active site residues, which are in close contact with bound substrate pyrroloquinoline quinone. It exhibits a stepwise process in which substrate binding leads to the generation of the closed protein conformation, with the latter playing a critical role in O2 binding and catalysis Klebsiella pneumoniae
1.3.3.11 physiological function pyrroloquinoline quinone, i.e. 4,5-dihydro-4,5-dioxo-1Hpyrrolo[2,3-f]quinoline-2,7,9-tricarboxylic acid, PQQ, synthesized by the enzyme, is a bacterial cofactor in numerous alcohol dehydrogenases including methanol dehydrogenase and glucose dehydrogenase Klebsiella pneumoniae