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 | 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 | 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 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 | 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 (Comment) | Organism |
---|---|
recombinant recombinant M subunit 8fold by ultracentrifugation, anion exchange chromatography, and gel filtration from Escherichia coli strain BL21(DE3) | Afipia carboxidovorans |
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 | Comment | Organism |
---|---|---|
Carbon monoxide dehydrogenase | - |
Afipia carboxidovorans |
CO dehydrogenase | - |
Afipia carboxidovorans |
molybdenum-containing carbon monoxide dehydrogenase | - |
Afipia carboxidovorans |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Afipia carboxidovorans |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.2 | - |
assay at | Afipia carboxidovorans |
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 | 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 |