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Identification and characterization of a bacterial hydrosulphide ion channel

Nature volume 483, pages 494–497 (2012)Cite this article

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Abstract

The hydrosulphide ion (HS−) and its undissociated form, hydrogen sulphide (H2S), which are believed to have been critical to the origin of life on Earth1, remain important in physiology and cellular signalling2. As a major metabolite in anaerobic bacterial growth, hydrogen sulphide is a product of both assimilatory and dissimilatory sulphate reduction2,3,4. These pathways can reduce various oxidized sulphur compounds including sulphate, sulphite and thiosulphate. The dissimilatory sulphate reduction pathway uses this molecule as the terminal electron acceptor for anaerobic respiration, in which process it produces excess amounts of H2S (ref. 4). The reduction of sulphite is a key intermediate step in all sulphate reduction pathways. In Clostridium and Salmonella, an inducible sulphite reductase is directly linked to the regeneration of NAD+, which has been suggested to have a role in energy production and growth, as well as in the detoxification of sulphite3. Above a certain concentration threshold, both H2S and HS− inhibit cell growth by binding the metal centres of enzymes and cytochrome oxidase5, necessitating a release mechanism for the export of this toxic metabolite from the cell5,6,7,8,9. Here we report the identification of a hydrosulphide ion channel in the pathogen Clostridium difficile through a combination of genetic, biochemical and functional approaches. The HS− channel is a member of the formate/nitrite transport family, in which about 50 hydrosulphide ion channels form a third subfamily alongside those for formate10,11 (FocA) and for nitrite12 (NirC). The hydrosulphide ion channel is permeable to formate and nitrite as well as to HS− ions. Such polyspecificity can be explained by the conserved ion selectivity filter observed in the channel’s crystal structure. The channel has a low open probability and is tightly regulated, to avoid decoupling of the membrane proton gradient.

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Figure 1: Genetic analyses and functional characterization of FNT3 and the asrABC operon.
Figure 2: Binding and transport activity of HSC in reconstituted proteoliposomes.
Figure 3: Structural and functional characterization of the ion permeation pathway.
Figure 4: Structural and functional characterization of possible gating mechanisms.

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Accession codes

Primary accessions

Protein Data Bank

Data deposits

The atomic coordinates and structure factors of HSC for high, medium and low pH have been deposited in the Protein Data Bank under accession codes 3TDO, 3TDR and 3TDP, respectively, and those of the Lys 16 Ser, Leu 82 Val, Thr 84 Ala, Lys 148 Glu and Phe 194 Ile mutants have been deposited under the codes 3TE2, 3TDX, 3TE1, 3TE0 and 3TDS, respectively.

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Acknowledgements

We are grateful the staff at beamlines X25 and X29 of the National Synchrotron Light Source at Brookhaven National Laboratory and at beamline 23ID of the Advanced Photon Source at Argonne National Laboratory for assistance in X-ray diffraction experiments. We thank A. B. Waight for suggesting the project; J. J. Marden for assistance with cloning of mutants; T. Neubert and S. Blais for mass spectrometry measurements; the 2010 CCP4 Workshop for assistance in processing diffraction data; and A. David, H. Jackson, N. K. Karpowich, J. J. Marden, R. L. Mancusso, Y. Pan and M. Zhou for discussions. This work was financially supported by the NIH (R01-GM093825, R01-DK073973, R01-MH083840 and U54-GM075026). B.K.C. was partly supported by an NIH Supplement Grant to Promote Diversity in Health-Related Research (R01-DK053973-08A1S1) and an NIH pre-doctoral fellowship (F31-AI086072).

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Authors and Affiliations

  1. The Helen L. and Martin S. Kimmel Center for Biology and Medicine at the Skirball Institute of Biomolecular Medicine, New York University School of Medicine, 540 First Avenue, New York, 10016, New York, USA

    Bryan K. Czyzewski & Da-Neng Wang

  2. Molecular Biophysics Graduate Program, New York University School of Medicine, 540 First Avenue, New York, 10016, New York, USA

    Bryan K. Czyzewski

  3. Department of Cell Biology, New York University School of Medicine, 540 First Avenue, New York, 10016, New York, USA

    Da-Neng Wang

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  1. Bryan K. Czyzewski
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B.K.C. did the experiments. B.K.C. and D.-N.W. wrote the manuscript.

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Correspondence to Da-Neng Wang.

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The authors declare no competing financial interests.

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Czyzewski, B., Wang, DN. Identification and characterization of a bacterial hydrosulphide ion channel. Nature 483, 494–497 (2012). https://doi.org/10.1038/nature10881

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