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Functional and structural characterization of the minimal Sec translocase of the hyperthermophile Thermotoga maritima

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Abstract

The genome of the hyperthermophilic bacterium Thermotoga maritima contains the genes that encode core subunits of the protein translocase, a complex consisting of the molecular motor SecA and the protein conducting pore SecYE. In addition, we identified an erroneous sequence in the genome encoding for a putative secG gene. The genes of the T. maritima translocase subunits were overexpressed in Escherichia coli and purified to homogeneity. T. maritima SecA showed a basal thermostable ATPase activity that was stimulated up to 4-fold by phospholipids with an optimum at 74°C. Membrane vesicles and proteoliposomes containing SecYE or SecYEG supported 2- to 4-fold stimulation of the precursor dependent SecA ATPase activity. Imaging of small two-dimensional crystals of the SecYE complex using electron microscopy showed square-shaped particles with a side-length of about 6 nm. These results demonstrate that in T. maritima a highly thermostable translocase complex is operational.

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Acknowledgements

M. G. Pretz and H. Remigy were supported by the MEMBMACS training network and funded by EU TMR contract HPRN-CT-2000-00075. S.V. Albers was supported by a VENI-grant from the Dutch Science Organization (NWO). We would like to thank to Dr. R Huber and Dr. M. Thomm from the University of Regensburg, Germany, for kindly providing us with T. maritima cells, and Dr. C. van der Does for valuable discussions.

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Author notes

  1. Jelto Swaving

    Present address: DSM Biologics, P.O. Box 454, 9700, Groningen, The Netherlands

Authors and Affiliations

  1. Department of Molecular Microbiology, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Kerklaan 30, 9751, Haren, The Netherlands

    Monika G. Pretz, Jelto Swaving, Sonja V. Albers, Victoria G. Garrido & Arnold J. M. Driessen

  2. Maurice E. Müller Institute for Microscopy, Biozentrum of the University of Basel, Kingelbergstrasse 70, 4056, Basel, Switzerland

    Hervé Remigy, Mohamed Chami & Andreas Engel

Authors
  1. Monika G. Pretz
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  2. Hervé Remigy
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  8. Arnold J. M. Driessen
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Corresponding author

Correspondence to Arnold J. M. Driessen.

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Communicated by G. Antranikian

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Pretz, M.G., Remigy, H., Swaving, J. et al. Functional and structural characterization of the minimal Sec translocase of the hyperthermophile Thermotoga maritima. Extremophiles 9, 307–316 (2005). https://doi.org/10.1007/s00792-005-0446-3

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  • DOI: https://doi.org/10.1007/s00792-005-0446-3

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