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Structure and function of polyamine-amino acid antiporters CadB and PotE in Escherichia coli

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Abstract

The structure and function of a cadaverine–lysine antiporter CadB and a putrescine–ornithine antiporter PotE in Escherichia coli were evaluated using model structures based on the crystal structure of AdiC, an agmatine–arginine antiporter, and the activities of various CadB and PotE mutants. The central cavity of CadB, containing the substrate binding site, was wider than that of PotE, mirroring the different sizes of cadaverine and putrescine. The size of the central cavity of CadB and PotE was dependent on the angle of transmembrane helix 6 (TM6) against the periplasm. Tyr73, Tyr89, Tyr90, Glu204, Tyr235, Asp303, and Tyr423 of CadB, and Cys62, Trp201, Glu207, Trp292, and Tyr425 of PotE were strongly involved in the antiport activities. In addition, Trp43, Tyr57, Tyr107, Tyr366, and Tyr368 of CadB were involved preferentially in cadaverine uptake at neutral pH, while only Tyr90 of PotE was involved preferentially in putrescine uptake. The results indicate that the central cavity of CadB consists of TMs 2, 3, 6, 7, 8, and 10, and that of PotE consists of TMs 2, 3, 6, and 8. These results also suggest that several amino acid residues are necessary for recognition of cadaverine in the periplasm because the level of cadaverine is much lower than that of putrescine in the periplasm at neutral pH. All the amino acid residues identified as being strongly involved in both the antiport and uptake activities were located on the surface of the transport path consisting of the central cavity and TM12.

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Abbreviations

AdiC:

Agmatine–arginine antiporter

CadB:

Cadaverine–lysine antiporter

PotE:

Putrescine–ornithine antiporter

APC superfamily:

Amino acid/polyamine/organocation superfamily

TM:

Transmembrane helix

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Acknowledgments

We thank Drs. A. J. Michael and K. Williams for their help in preparing this manuscript.

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Authors and Affiliations

  1. Faculty of Pharmacy, Chiba Institute of Science, Choshi, Chiba, Japan

    Hideyuki Tomitori & Keiko Kashiwagi

  2. Amine Pharma Research Institute, Innovation Plaza at Chiba University, Chiba, Chiba, Japan

    Kazuei Igarashi

  3. Graduate School of Pharmaceutical Sciences, Chiba University, 1-8-1, Inohana, Chuo-ku, Chiba, Chiba, 260-8675, Japan

    Kazuei Igarashi

Authors
  1. Hideyuki Tomitori
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  2. Keiko Kashiwagi
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  3. Kazuei Igarashi
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Correspondence to Kazuei Igarashi.

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Tomitori, H., Kashiwagi, K. & Igarashi, K. Structure and function of polyamine-amino acid antiporters CadB and PotE in Escherichia coli . Amino Acids 42, 733–740 (2012). https://doi.org/10.1007/s00726-011-0989-9

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  • DOI: https://doi.org/10.1007/s00726-011-0989-9

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