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NMR resonance assignments of the lantibiotic immunity protein NisI from Lactococcus lactis

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Abstract

The lantibiotic nisin is a small antimicrobial peptide which acts against a wide range of Gram-positive bacteria. Nisin-producing Lactococcus lactis strains express four genes for self-protection against their own antimicrobial compound. This immunity system consists of the lipoprotein NisI and the ABC transporter NisFEG. NisI is attached to the outside of the cytoplasmic membrane via a covalently linked diacylglycerol anchor. Both the lipoprotein and the ABC transporter are needed for full immunity but the exact immunity mechanism is still unclear. To gain insights into the highly specific immunity mechanism of nisin producing strains on a structural level we present here the backbone resonance assignment of NisI (25.8 kDa) as well as the virtually complete 1H,15N,13C chemical shift assignments for the isolated 12.7 kDa N-terminal and 14.6 kDa C-terminal domains of NisI.

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References

  • Bierbaum G, Sahl HG (2009) Lantibiotics: mode of action, biosynthesis and bioengineering. Curr Pharm Biotechnol 10:2–18

    Article  Google Scholar 

  • Breukink E, Wiedemann I, van Kraaij C, Kuipers OP, Sahl H, de Kruijff B (1999) Use of the cell wall precursor lipid II by a pore-forming peptide antibiotic. Science 286:2361–2364

    Article  Google Scholar 

  • Brötz H, Josten M, Wiedemann I, Schneider U, Gotz F, Bierbaum G, Sahl HG (1988) Role of lipid-bound peptidoglycan precursors in the formation of pores by nisin, epidermin and other lantibiotics. Mol Microbiol 30:317–327

    Article  Google Scholar 

  • Christ NA, Duchardt-Ferner E, Düsterhus S, Kötter P, Entian KD, Wöhnert J (2012a) NMR resonance assignment of the autoimmunity protein SpaI from Bacillus subtilis ATCC 6633. Biomol NMR Assign 6:9–13

    Article  Google Scholar 

  • Christ NA, Bochmann S, Gottstein D, Duchardt-Ferner E, Hellmich UA, Düsterhus S, Kütter P, Güntert P, Entian K, Wöhnert J (2012b) The first structure of a lantibiotic immunity protein, SpaI from Bacillus subtilis, reveals a novel fold. J Biol Chem 287:35286–35298

    Article  Google Scholar 

  • Farrow NA, Muhandiram R, Singer AU, Pascal SM, Kay CM, Gish G et al (1994) Backbone dynamics of a free and phosphopeptide-complexed Src homology 2 domain studied by 15 N NMR relaxation. Biochemistry 33:5984–6003

    Article  Google Scholar 

  • Hasper HE, Kramer NE, Smith JL, Hillman JD, Zachariah C, Kuipers OP, de Kruijff B, Breukink E (2006) An alternative bactericidal mechanism of action for lantibiotic peptides that target lipid II. Science 313:1636–1637

    Article  ADS  Google Scholar 

  • Hsu SD, Breukink E, Tischenko E, Lutters MAG, de Kruijff B, Kaptein R, Bonvin AMJJ, van Nuland NAJ (2004) The nisin–lipid II complex reveals a pyrophosphate cage that provides a blueprint for novel antibiotics. Nat Struct Mol Biol 11:963–967

    Article  Google Scholar 

  • Keller R (2004) The Computer Aided Resonance Tutorial. CANTINA Verlag, Goldau

    Google Scholar 

  • Klein C, Entian KD (1994) Genes involved in self-protection against the lantibiotic subtilin produced by Bacillus subtilis ATCC 6633. Appl Environ Microbiol 60:2793–2801

    Google Scholar 

  • Koponen O, Takala TM, Saarela U, Qiao M, Saris PEJ (2004) Distribution of the NisI immunity protein and enhancement of nisin activity by the lipid-free NisI. FEMS Microbiol Lett 231:85–90

    Article  Google Scholar 

  • Kuipers OP, Beerthuyzen MM, Siezen RJ, De Vos WM (1993) Characterization of the nisin gene cluster nisABTCIPR of Lactococcus lactis. Requirement of expression of the nisA and nisI genes for development of immunity. Eur J Biochem 216:281–291

    Article  Google Scholar 

  • Markley JL, Bax A, Arata Y, Hilbers CW, Kaptein R, Sykes BD, Wright PE, Wüthrich 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 12:1–23

    Article  Google Scholar 

  • Muchmore DC, McIntosh LP, Russell CB, Anderson de, Dahlquist FW (1989) Expression and nitrogen-15 labeling of proteins for proton and nitrogen-15 nuclear magnetic resonance. Methods Enzymol 177:44–73

  • Qiao M, Immonen T, Koponen O, Saris PE (1995) The cellular location and effect on nisin immunity of the NisI protein from Lactococcus lactis N8 expressed in Escherichia coli and L. lactis. FEMS Microbiol Lett 131:75–80

    Article  Google Scholar 

  • Rogers LA (1928) The inhibiting effect of Streptococcus lactis on Lactobacillus bulgaricus. J Bacteriol 16:321–325

  • Salzmann M, Pervushin K, Wider G, Senn H, Wüthrich K (1998) TROSY in triple-resonance experiments: new perspectives for sequential NMR assignment of large proteins. Proc Natl Acad Sci USA 95:13585–13590

    Article  ADS  Google Scholar 

  • Sattler M, Schleucher J, Griesinger C (1999) Heteronuclear multidimensional NMR experiments for the structure determination of proteins in solution employing pulsed field gradients. Prog Nucl Magn Reson Spectrosc 34:93–158

    Article  Google Scholar 

  • Schnell N, Entian KD, Schneider U, Götz F, Zähner H, Kellner R, Jung G (1988) Prepeptide sequence of epidermin, a ribosomally synthesized antibiotic with four sulphide-rings. Nature 333:276–278

    Article  ADS  Google Scholar 

  • Shen Y, Bax A (2010) Prediction of Xaa-Pro peptide bond conformation from sequence and chemical shifts. J Biomol NMR 46:199–204

    Article  Google Scholar 

  • Shen Y, Bax A (2013) Protein backbone and sidechain torsion angles predicted from NMR  chemical shifts using artificial neural networks. J Biomol NMR 56:227–241

  • Siegers K, Entian KD (1995) Genes involved in immunity to the lantibiotic nisin produced by Lactococcus lactis 6F3. Appl Environ Microbiol 61:1082–1089

    Google Scholar 

  • Stein T, Heinzmann S, Solovieva I, Entian K (2003) Function of Lactococcus lactis nisin immunity genes nisI and nisFEG after coordinated expression in the surrogate host Bacillus subtilis. J Biol Chem 278:89–94

    Article  Google Scholar 

  • Takala TM, Koponen O, Qiao M, Saris PE (2004) Lipid-free NisI: interaction with nisin and contribution to nisin immunity via secretion. FEMS Microbiol Lett 237:171–177

    Article  Google Scholar 

  • Vranken WF, Boucher W, Stevens TJ, Fogh RH, Pajon A, Llinas M et al (2005) The CCPN data model for NMR spectroscopy: development of a software pipeline. Proteins 59:687–696

    Article  Google Scholar 

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Acknowledgments

We thank Prof. Volker Dötsch for the kind gift of the TEV protease construct.This project was supported by the DFG (Wo 901/4-1 to J.W. and En 134/11-1 to K.-D.E.), an Aventis Foundation professorship (to J.W.) and the Center of Biomolecular Magnetic Resonance (BMRZ) at the Goethe University Frankfurt am Main.

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Correspondence to Jens Wöhnert.

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Hacker, C., Christ, N.A., Duchardt-Ferner, E. et al. NMR resonance assignments of the lantibiotic immunity protein NisI from Lactococcus lactis . Biomol NMR Assign 9, 293–297 (2015). https://doi.org/10.1007/s12104-015-9595-1

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