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Literature summary for 1.2.5.3 extracted from

  • Gremer, L.; Kellner, S.; Dobbek, H.; Huber, R.; Meyer, O.
    Binding of flavin adenine dinucleotide to molybdenum-containing carbon monoxide dehydrogenase from Oligotropha carboxidovorans. Structural and functional analysis of a carbon monoxide dehydrogenase species in which the native flavoprotein has been replaced by its recombinant counterpart produced in Escherichia coli (2000), J. Biol. Chem., 275, 1864-1872.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
genes coxS, coxM, and coxL, recombinant expression of FAD-reconstituted enzyme and of monomeric deflavo medium subunit in Escherichia coli strain BL21(DE3), subcloning in Escherichia coli strain DH5alpha Afipia carboxidovorans

Protein Variants

Protein Variants Comment Organism
additional information the flavoprotein can be removed from CO dehydrogenase by dissociation with sodium dodecylsulfate, the resulting M(LS)2- or (LS)2-structured CO dehydrogenase species can be reconstituted with the recombinant apoflavoprotein produced in Escherichia coli. Binding of FAD to the reconstituted deflavo (LMS)2 species occurs with second-order kinetics and high affinity. Same fold and binding of the flavoprotein as in wild-type CO dehydrogenase, whereas the S-selanylcysteine 388 in the active-site loop on the molybdoprotein is disordered. The structural changes related to heterotrimeric complex formation or FAD binding are transmitted to the iron-sulfur protein, structural and functional analysis of FAD binding in CO dehydrogenase Afipia carboxidovorans

Metals/Ions

Metals/Ions Comment Organism Structure
Fe2+ in two [2Fe-2S] clusters Afipia carboxidovorans
Molybdenum in the molybdoprotein Afipia carboxidovorans
selenium an S-selanylcysteine-containing large subunit Afipia carboxidovorans
[2Fe-2S] cluster the type II 2Fe:2S center is identified in the N-terminal domain and the type I center in the C-terminal domain of the iron-sulfur protein Afipia carboxidovorans

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
17800
-
(alphabetagamma)2, 2 * 88700, large subunit, + 2 * 30200, medoum subunit, + 2 * 17800, small subunit, SDS-PAGE Afipia carboxidovorans
30200
-
(alphabetagamma)2, 2 * 88700, large subunit, + 2 * 30200, medoum subunit, + 2 * 17800, small subunit, SDS-PAGE Afipia carboxidovorans
41000
-
recombinant monomeric deflavo medium subunit, gel filtration Afipia carboxidovorans
88700
-
(alphabetagamma)2, 2 * 88700, large subunit, + 2 * 30200, medoum subunit, + 2 * 17800, small subunit, SDS-PAGE Afipia carboxidovorans

Organism

Organism UniProt Comment Textmining
Afipia carboxidovorans P19919 and P19920 and P19921 genes coxL, coxM, and coxS; formerly Pseudomonas carboxydovorans strain OM5, genes coxS, coxM, and coxL
-
Afipia carboxidovorans DSM 1227 P19919 and P19920 and P19921 genes coxL, coxM, and coxS; formerly Pseudomonas carboxydovorans strain OM5, genes coxS, coxM, and coxL
-

Purification (Commentary)

Purification (Comment) Organism
recombinant recombinant M subunit 8fold by ultracentrifugation, anion exchange chromatography, and gel filtration from Escherichia coli strain BL21(DE3) Afipia carboxidovorans

Subunits

Subunits Comment Organism
heterohexamer (alphabetagamma)2, 2 * 88700, large subunit, + 2 * 30200, medoum subunit, + 2 * 17800, small subunit, SDS-PAGE Afipia carboxidovorans
More the enzyme is an S-selanylcysteine-containing 88.7-kDa molybdoprotein, a 17.8-kDa iron-sulfur protein, and a 30.2-kDa flavoprotein in a (LMS)2 subunit structure Afipia carboxidovorans

Synonyms

Synonyms Comment Organism
Carbon monoxide dehydrogenase
-
Afipia carboxidovorans
CO dehydrogenase
-
Afipia carboxidovorans
molybdenum-containing carbon monoxide dehydrogenase
-
Afipia carboxidovorans

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
assay at Afipia carboxidovorans

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.2
-
assay at Afipia carboxidovorans

Cofactor

Cofactor Comment Organism Structure
FAD FAD is bound in the medium subunit. The flavoprotein can be removed from CO dehydrogenase by dissociation with sodium dodecylsulfate, the resulting M(LS)2- or (LS)2-structured CO dehydrogenase species can be reconstituted with the recombinant apoflavoprotein produced in Escherichia coli, structural and functional analysis of FAD binding in CO dehydrogenase Afipia carboxidovorans
molybdopterin cofactor the molybdoprotein of CO dehydrogenase carries the molybdopterin cytosine dinucleotide (MCD)1-type of molybdenum cofactor and the unique active-site loop Gly383-Val-Ala-Tyr-Arg-Cys-Ser-Phe-Arg391, which positions the catalytically essential S-selanylcysteine 388 in a distance of 3.7 A to the molybdenum ion Afipia carboxidovorans
additional information an S-selanylcysteine-containing 88.7-kDa molybdoprotein, a 17.8-kDa iron-sulfur protein, and a 30.2-kDa flavoprotein in a (LMS)2 subunit structure Afipia carboxidovorans

General Information

General Information Comment Organism
evolution CO dehydrogenase is a prototype of the molybdenum hydroxylase sequence family Afipia carboxidovorans
additional information the formation of the heterotrimeric complex composed of the apoflavoprotein, the molybdoprotein, and the iron-sulfur protein involves structural changes that translate into the conversion of the apoflavoprotein from non-FAD binding to FAD binding Afipia carboxidovorans