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NMR resonance assignment of the autoimmunity protein SpaI from Bacillus subtilis ATCC 6633

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Abstract

Bacillus subtilis ATCC 6633 produces the lipid II targeting lantibiotic subtilin. For self-protection these gram-positive bacteria express a cluster of four self-immunity proteins named SpaIFEG. SpaI is a 16.8 kDa lipoprotein which is attached to the outside of the cytoplasmic membrane via a covalently linked diacylglycerol anchor. Together with the ABC-transporter SpaFEG, SpaI protects the membrane from subtilin insertion and there is evidence for a direct interaction of SpaI with subtilin. As a prerequisite for further structural studies of SpaI and the SpaI/subtilin complex we report here the full 1H, 15N, 13C chemical shift assignment for a stable 14.9 kDa C-terminal fragment of SpaI.

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Acknowledgments

We are grateful to Dr. Christian Richter for help with the NMR experiments and to Sophie Bochmann for helpful discussions. We thank Prof. Volker Dötsch for the kind gift of the TEV-protease construct and Björn Meyer and Prof. Michael Karas for the MALDI-MS analysis of the C-terminal SpaI fragment. The eNMR project (European FP7 e-Infrastructure grant, contract no. 213010, www.enmr.eu), supported by the national GRID Initiatives of Italy, Germany and the Dutch BiG Grid project (Netherlands Organization for Scientific Research) is acknowledged for the use of web portals, computing and storage facilities. This project was supported by an Aventis Foundation professorship (to J.W.), the Center of Biomolecular Magnetic Resonance (BMRZ), the Cluster of Excellence “Macromolecular Complexes” and the Förderfonds of the Goethe University Frankfurt am Main.

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Authors and Affiliations

  1. Institut für Molekulare Biowissenschaften, Johann-Wolfgang-Goethe-Universität Frankfurt/M., Max-von-Laue-Str. 9, 60438, Frankfurt, Germany

    Nina Alexandra Christ, Elke Duchardt-Ferner, Stefanie Düsterhus, Peter Kötter, Karl-Dieter Entian & Jens Wöhnert

  2. Center of Biomolecular Magnetic Resonance (BMRZ), Johann-Wolfgang-Goethe-Universität Frankfurt/M., Max-von-Laue-Str. 9, 60438, Frankfurt, Germany

    Nina Alexandra Christ, Elke Duchardt-Ferner & Jens Wöhnert

  3. Cluster of Excellence “Macromolecular Complexes”, Johann-Wolfgang-Goethe-Universität, Max-von-Laue-Str. 9, 60438, Frankfurt, Germany

    Karl-Dieter Entian & Jens Wöhnert

Authors
  1. Nina Alexandra Christ
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  2. Elke Duchardt-Ferner
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  3. Stefanie Düsterhus
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  4. Peter Kötter
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  5. Karl-Dieter Entian
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  6. Jens Wöhnert
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Correspondence to Jens Wöhnert.

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Christ, N.A., Duchardt-Ferner, E., Düsterhus, S. et al. NMR resonance assignment of the autoimmunity protein SpaI from Bacillus subtilis ATCC 6633. Biomol NMR Assign 6, 9–13 (2012). https://doi.org/10.1007/s12104-011-9314-5

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