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1H, 13C, and 15N backbone, side-chain, and heme chemical shift assignments for oxidized and reduced forms of the monoheme c-type cytochrome ApcA isolated from the acidophilic metal-reducing bacterium Acidiphilium cryptum

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Abstract

We report the 1H, 13C, and 15N chemical shift assignments of both oxidized and reduced forms of an abundant periplasmic c-type cytochrome, designated ApcA, isolated from the acidophilic gram-negative facultatively anaerobic metal-reducing alphaproteobacterium Acidiphilium cryptum. These resonance assignments prove that ApcA is a monoheme cytochrome c 2 and the product of the Acry_2099 gene. An absence of resonance peaks in the NMR spectra for the 21N-terminal residues suggests that a predicted N-terminal signal sequence is cleaved. We also describe the preparation and purification of the protein in labeled form from laboratory cultures of A. cryptum growing on 13C- and 15N- labeled substrates.

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Acknowledgments

600 and 750 MHz NMR spectra were acquired in the Environmental Molecular Sciences Laboratory (EMSL), a national scientific user facility sponsored by the Department of Energy’s Office of Biological and Environmental Research and located at Pacific Northwest National Laboratory. Research was supported by the Department of Energy (Grant DE-FG02-06ER15824 to TSM) and the National Science Foundation (Grant 0434023 to TSM).

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Authors and Affiliations

  1. Biological Sciences Division, Pacific Northwest National Laboratory, Richland, WA, 99352, USA

    John R. Cort

  2. Department of Biological Sciences, Idaho State University, Pocatello, ID, 83204, USA

    Michael W. Swenson & Timothy S. Magnuson

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  1. John R. Cort
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  2. Michael W. Swenson
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  3. Timothy S. Magnuson
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Correspondence to John R. Cort.

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Fig. S1

600 MHz 1H–13C HSQC spectra recorded in D2O showing the aliphatic regions for oxidized (top) and reduced (bottom) ApcA at pH 5.0. Spectra were recorded at 293 K with 192 or 256 complex points in the indirect dimension and 1024 complex points acquired in the direct dimension. The spectral width of the 13C dimension was 80 ppm in both spectra. (EPS 7622 kb)

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Cort, J.R., Swenson, M.W. & Magnuson, T.S. 1H, 13C, and 15N backbone, side-chain, and heme chemical shift assignments for oxidized and reduced forms of the monoheme c-type cytochrome ApcA isolated from the acidophilic metal-reducing bacterium Acidiphilium cryptum . Biomol NMR Assign 5, 89–92 (2011). https://doi.org/10.1007/s12104-010-9274-1

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  • DOI: https://doi.org/10.1007/s12104-010-9274-1

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