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Electron transfer in Paracoccus denitrificans with the modified fbc operon

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Abstract

Membrane fragments of two mutant strains of Paracoccus denitrificans genetically modified in the bc 1 complex have been studied for comparison of enzymic activities of succinate-cytochrome-c reductase and its components, viz. succinate dehydrogenase (Complex II) and ubiquinol-cytochrome-c reductase (Complex III) and their response to changes in concentration of succinate, cytochrome c, ionic strength, pH, temperature and sensitivity to antimycin A. The mutants synthesized and assembled the b and c hemes in the ratio characteristic for the wild type strain. The mutant strain M 71 expressing the truncated copy of cytochrome c 1 (devoid of a stretch of 150 mainly acidic amino acids) was less sensitive to increasing concentration of cytochrome c and changes in ionic strength of the medium, but maintained the original affinity to succinate and sensitivity to antimycin A. The mutant strain M 36 with an overexpressed bc 1 content showed the highest response to changes in ionic strength and physical parameters, exhibited the lowest turnover number values with succinate-cytochrome-c reductase, but positively affected the succinate dehydrogenase. In view of the interaction of the redox components in native membranes the functional analyses of separated Complexes II and III should be regarded with caution.

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Abbreviations

cyt:

cytochrome

DCIP:

2,6-dichloroindophenol

ISP(s):

iron-sulfur cluster(s) (Fe-S protein)

k (1/s):

first-order rate constant

k cat :

turnover number value(s)

M 36:

mutant strain G440/pEG436

M 71:

mutant strain G440/pEG471

CYO:

cytochrome-c oxidase (EC 1.9.3.1)

QCR:

ubiquinol-cytochrome-c reductase (EC 1.10.2.2)

PMS:

phenazine methosulfate

Q:

ubiquinone

QH2 :

ubiquinol

ROS:

reactive oxygen species

RSD:

relative standard deviation

Suc:

succinate

WT:

wild-type (strain)

SCR:

succinate-cytochrome-c reductase (see the text)

SDH:

succinate dehydrogenase (EC 1.3.99.1)

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Authors and Affiliations

  1. Institute of Biochemistry, Faculty of Science, Masaryk University, 625 00, Brno, Czechia

    V. Dadák, J. Dudák & P. Zbořil

  2. Department of Biochemistry, Faculty of Science, Masaryk University, 611 37, Brno, Czech Republic

    V. Dadák

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  1. V. Dadák
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  2. J. Dudák
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  3. P. Zbořil
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Correspondence to V. Dadák.

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Dadák, V., Dudák, J. & Zbořil, P. Electron transfer in Paracoccus denitrificans with the modified fbc operon. Folia Microbiol 54, 475–482 (2009). https://doi.org/10.1007/s12223-009-0067-9

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  • DOI: https://doi.org/10.1007/s12223-009-0067-9

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