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Redox properties of the Rhodobacter sphaeroides transcriptional regulatory proteins PpsR and AppA

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Abstract

Redox properties of the photosynthetic gene repressor PpsR and the blue-light photoreceptor/antirepressor AppA from Rhodobacter sphaeroides have been characterized. Redox titrations of PpsR reveal the presence of a two-electron couple, with an E m value of −320 mV at pH 7.0, which is likely to arise from the reversible conversion of two cysteine thiols to a disulfide. This E m value is very much more negative than the E m = −180 mV value measured previously at pH 7.0 for the disulfide/dithiol couple in CrtJ, the homolog for PpsR in the closely related bacterium Rhodobacter capsulatus. AppA, a flavin-containing blue-light receptor that is also involved in the regulation of gene expression in R. sphaeroides, contains multiple cysteines in its C-terminal region, two of which function as a redox-active dithiol/disulfide couple with an E m value of −325 mV at pH 7.0 in the dark. Titrations of this dithiol/disulfide couple in illuminated samples of AppA indicate that the E m value of this disulfide/dithiol couple is −315 mV at pH 7.0, identical to the value obtained for AppA in the dark within the combined experimental uncertainties of the two measurements. The E m values of AppA and PpsR demonstrate that these proteins are thermodynamically capable of electron transfer for their activity as an anti-repressor/repressor in R. sphaeroides.

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Abbreviations

mV:

millivolt

DTNB:

5,5′-dithiolbis(2-nitrobenzoic acid)

mBBr:

monobromobimane

DTT:

dithiothreitol

GSH:

reduced glutathione

GSSG:

oxidized glutathione

E m :

midpoint potential

E h :

ambient potential

AMS:

4-acetamido-4′-maleimidylstibene-2,2′-disulfonic acid

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Acknowledgments

This work was funded by grants from the Robert A. Welch Foundation (D-0710 to DBK) and from National Institutes of Health (GM040941 to CEB).

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Authors and Affiliations

  1. Department of Chemistry and Biochemistry, Texas Tech University, Lubbock, TX, 79409-1061, USA

    S.-K. Kim, J. T. Mason & D. B. Knaff

  2. Institute for Biotechnology and Genomics, Texas Tech University, Lubbock, TX, 79409-1061, USA

    D. B. Knaff

  3. Department of Biology, Indiana University, 915 E. 3rd St., Myers Hall, Bloomington, IN, 47405, USA

    C. E. Bauer & A. T. Setterdahl

Authors
  1. S.-K. Kim
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  2. J. T. Mason
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  3. D. B. Knaff
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  4. C. E. Bauer
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  5. A. T. Setterdahl
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Correspondence to A. T. Setterdahl.

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Kim, SK., Mason, J.T., Knaff, D.B. et al. Redox properties of the Rhodobacter sphaeroides transcriptional regulatory proteins PpsR and AppA. Photosynth Res 89, 89–98 (2006). https://doi.org/10.1007/s11120-006-9086-4

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  • DOI: https://doi.org/10.1007/s11120-006-9086-4

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