NMR resonance assignment of the autoimmunity protein SpaI from Bacillus subtilis ATCC 6633
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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.
KeywordsNMR-assignments Triple resonance experiments SpaI Self-immunity Lantibiotic Subtilin
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.
- Engelke G, Gutowski-Eckel Z, Kiesau P, Siegers K, Hammelmann M, Entian KD (1994) Regulation of nisin biosynthesis and immunity in Lactococcus lactis 6F3. Appl Environ Microbiol 60:814–825Google Scholar
- Gross E (1975) Subtilin and nisin: the chemistry and biology of peptides with alpha, beta-unsaturated amino acids, p. 31–42. In: R.Walter and J. Meienhofer (ed.), Peptides: chemistry, structure and biology: proceedings of the fourth American Peptide Symposium. Ann Arbor Sciecne, Ann Arbor, MI., pp. 31–42Google Scholar
- Keller R (2004) The computer aided resonance tutorial. CANTINA Verlag, GoldauGoogle Scholar
- Klein C, Entian KD (1994) Genes involved in self-protection against the lantibiotic subtilin produced by Bacillus subtilis ATCC 6633. Appl Environ Microbiol 60:2793–2801Google Scholar
- Markley JL, Bax A, Arata Y, Hilbers CW, Kaptein R, Sykes BD, Wright PE, Wuthrich K (1998) Recommendations for the presentation of NMR structures of proteins and nucleic acids. IUPAC-IUBMB-IUPAB inter-union task group on the standardization of data bases of protein and nucleic acid structures determined by NMR spectroscopy. J Biomol NMR 12:1–23CrossRefGoogle Scholar