Expression, purification, and characterization of the CuA–cytochrome c domain from subunit II of the Bacillus subtilis cytochrome caa3 complex in Escherichia coli

Andrews, Diann; Mattatall, Neil R.; Arnold, Danielle E.; Hill, Bruce C.

doi:10.1016/j.pep.2004.11.009

Cited by 8 publications

(6 citation statements)

References 25 publications

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“…Also unique to this strain are three proteins involved in the biogenesis of cytochrome caa3 oxidase. Cytochrome caa 3 oxidase is the major oxidase involved in the last stages of the respiratory electron transport chain in B. subtilis grown under aerobic conditions, transferring electrons from the cytochrome c in the respiratory chain to the terminal electron acceptor, oxygen [ 24 , 25 ]. Deletion of the structural genes for cytochrome caa3 oxidase in B. subtilis showed that this enzyme is not essential for growth [ 26 ].…”

Section: Resultsmentioning

confidence: 99%

Comparative genomic analysis of Parageobacillus thermoglucosidasius strains with distinct hydrogenogenic capacities

et al. 2018

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BackgroundThe facultatively anaerobic thermophile Parageobacillus thermoglucosidasius produces hydrogen gas (H2) by coupling CO oxidation to proton reduction in the water-gas shift (WGS) reaction via a carbon monoxide dehydrogenase–hydrogenase enzyme complex. Although little is known about the hydrogenogenic capacities of different strains of this species, these organisms offer a potentially viable process for the synthesis of this alternative energy source.ResultsThe WGS-catalyzed H2 production capacities of four distinct P. thermoglucosidasius strains were determined by cultivation and gas analysis. Three strains (DSM 2542T, DSM 2543 and DSM 6285) were hydrogenogenic, while the fourth strain (DSM 21625) was not. Furthermore, in one strain (DSM 6285) H2 production commenced earlier in the cultivation than the other hydrogenogenic strains. Comparative genomic analysis of the four strains identified extensive differences in the protein complement encoded on the genomes, some of which are postulated to contribute to the different hydrogenogenic capacities of the strains. Furthermore, polymorphisms and deletions in the CODH-NiFe hydrogenase loci may also contribute towards this variable phenotype.ConclusionsDisparities in the hydrogenogenic capacities of different P. thermoglucosidasius strains were identified, which may be correlated to variability in their global proteomes and genetic differences in their CODH-NiFe hydrogenase loci. The data from this study may contribute towards an improved understanding of WGS-catalysed hydrogenogenesis by P. thermoglucosidasius.Electronic supplementary materialThe online version of this article (10.1186/s12864-018-5302-9) contains supplementary material, which is available to authorized users.

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Section: Resultsmentioning

confidence: 99%

Comparative genomic analysis of Parageobacillus thermoglucosidasius strains with distinct hydrogenogenic capacities

et al. 2018

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“…The plasmid pMA10 containing the gene of Cu A protein inserted to NcoI and BamHI restriction sites was a kind gift from Prof. B. Ludwig, Biozentrum, Goethe Universität, Frankfurt, Germany. The expression of the soluble Cu A domain was done by a procedure based on previous reports, protein was purified by chromatography on a DEAE Sepharose column using 50 mM Tris-HCl pH 6.5 buffer as eluent [37]. Purified Cu A was eluted in the fractions with Rz (A 280 /A 477 ) value more than 5.…”

Section: Methodsmentioning

confidence: 99%

Direct electrochemistry of dinuclear CuA fragment from cytochrome c oxidase of Thermus thermophilus at surfactant modified glassy carbon electrode

Rajbongshi

Das

Mazumdar

2010

Electrochimica Acta

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“…The fusion proteins were cleaved using thrombin, and thrombin was removed when the samples were passed over benzamidinelinked Sepharose. The purity of the BsSco preparations were assessed by denaturing polyacylamide electrophoresis as described previously (18).…”

Section: Methodsmentioning

confidence: 99%

Probing the Kinetics and Thermodynamics of Copper(II) Binding toBacillus subtilisSco, a Protein Involved in the Assembly of the Cu_ACenter of CytochromecOxidase

Cawthorn¹,

Poulsen²,

Davidson³

et al. 2009

Biochemistry

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BsSco is a member of the Sco protein family involved in the assembly of the Cu(A) center within cytochrome c oxidase. BsSco forms a complex with Cu(II) that has properties consistent with dithiolate ligation. Stopped-flow UV-visible absorbance and fluorescence coupled with multiwavelength analysis reveal biphasic binding kinetics between BsSco and Cu(II). An initial species appears with absorbance centered at 382 nm at a copper concentration-dependent rate (2.9 x 10(4) M(-1) s(-1)). The initial species decays at a first-order rate (1.5 s(-1)) to the equilibrium form with a maximum at 352 nm. Formation of the BsSco-Cu(II) complex is accompanied by quenching of protein fluorescence. The copper concentration-dependent phase gives 70% of the total quenching, while the final 30% develops during the second phase of the absorbance change. The pH dependence of copper binding shows that the copper-dependent rate increases by 50-fold as the pH decreases from 8.5 to 5.5 with an apparent pK(a) of 6.7. The slower phase rate is independent of pH. Comparison of circular dichroism spectra between apo-BsSco and the BsSco-Cu(II) complex reveals a small change in the UV region consistent with a subtle conformational change upon copper binding. There is formation of a distinctive visible CD spectrum in the BsSco-Cu(II) complex. A model is presented in which the kinetic and thermodynamic stability of the BsSco-Cu(II) complex results from a two-step mechanism. Release of copper would be facilitated in the intermediate form of BsSco, and attaining such a low-Cu(II) affinity state may be important for BsSco's function in Cu(A) assembly.

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Expression, purification, and characterization of the CuA–cytochrome c domain from subunit II of the Bacillus subtilis cytochrome caa3 complex in Escherichia coli

Cited by 8 publications

References 25 publications

Comparative genomic analysis of Parageobacillus thermoglucosidasius strains with distinct hydrogenogenic capacities

Comparative genomic analysis of Parageobacillus thermoglucosidasius strains with distinct hydrogenogenic capacities

Direct electrochemistry of dinuclear CuA fragment from cytochrome c oxidase of Thermus thermophilus at surfactant modified glassy carbon electrode

Probing the Kinetics and Thermodynamics of Copper(II) Binding toBacillus subtilisSco, a Protein Involved in the Assembly of the Cu_ACenter of CytochromecOxidase

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Expression, purification, and characterization of the CuA–cytochrome c domain from subunit II of the Bacillus subtilis cytochrome caa3 complex in Escherichia coli

Cited by 8 publications

References 25 publications

Comparative genomic analysis of Parageobacillus thermoglucosidasius strains with distinct hydrogenogenic capacities

Comparative genomic analysis of Parageobacillus thermoglucosidasius strains with distinct hydrogenogenic capacities

Direct electrochemistry of dinuclear CuA fragment from cytochrome c oxidase of Thermus thermophilus at surfactant modified glassy carbon electrode

Probing the Kinetics and Thermodynamics of Copper(II) Binding toBacillus subtilisSco, a Protein Involved in the Assembly of the CuACenter of CytochromecOxidase

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Probing the Kinetics and Thermodynamics of Copper(II) Binding toBacillus subtilisSco, a Protein Involved in the Assembly of the Cu_ACenter of CytochromecOxidase