Abstract
Corynebacterium glutamicum is used in microbial biotechnology for the production of amino acids, e.g., glutamate and lysine. Excretion of glutamate into the surrounding medium under production conditions is mediated by MscCG, an MscS-type mechanosensitive channel. In difference to most other MscS-type channel proteins, MscCG carries, in addition to the N-terminal pore domain, a long C-terminal domain that amounts to about half of the size of the protein and harbors an additional transmembrane segment. Here we study the impact of the C-terminal domain on both functions of MscCG as mechanosensitive channel and as glutamate exporter. Sequential truncations of the C-terminal domain were applied, as well as deletion of particular subdomains, replacement of these segments by other amino acid sequences, and sequence randomization. Several parameters of cell physiology and bioenergetics of the obtained mutants related to both glutamate excretion and response to osmotic stress were quantified. All three subdomains of the C-terminal domain, i.e., the periplasmic loop, the fourth transmembrane segment, and the cytoplasmic loop, proved to be of core significance for MscCG function, in particular for glutamate excretion.
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Acknowledgments
We gratefully acknowledge valuable discussions with Boris Martinac, Takeshi Nomura, Andrew Battle, and Yoshitaka Nakayama (Victor Chang Cardiac Research Institute, Sydney, Australia) within the joint project on MscCG. We would like to thank Anja Wittmann for excellent technical assistance.
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Special issue: Biophysics of Mechanotransduction.
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Becker, M., Krämer, R. MscCG from Corynebacterium glutamicum: functional significance of the C-terminal domain. Eur Biophys J 44, 577–588 (2015). https://doi.org/10.1007/s00249-015-1041-x
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DOI: https://doi.org/10.1007/s00249-015-1041-x