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Action of the multifunctional peptide BP100 on native biomembranes examined by solid-state NMR

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Abstract

Membrane composition is a key factor that regulates the destructive activity of antimicrobial peptides and the non-leaky permeation of cell penetrating peptides in vivo. Hence, the choice of model membrane is a crucial aspect in NMR studies and should reflect the biological situation as closely as possible. Here, we explore the structure and dynamics of the short multifunctional peptide BP100 using a multinuclear solid-state NMR approach. The membrane alignment and mobility of this 11 amino acid peptide was studied in various synthetic lipid bilayers with different net charge, fluidity, and thickness, as well as in native biomembranes harvested from prokaryotic and eukaryotic cells. 19F-NMR provided the high sensitivity and lack of natural abundance background that are necessary to observe a labelled peptide even in protoplast membranes from Micrococcus luteus and in erythrocyte ghosts. Six selectively 19F-labeled BP100 analogues gave remarkably similar spectra in all of the macroscopically oriented membrane systems, which were studied under quasi-native conditions of ambient temperature and full hydration. This similarity suggests that BP100 has the same surface-bound helical structure and high mobility in the different biomembranes and model membranes alike, independent of charge, thickness or cholesterol content of the system. 31P-NMR spectra of the phospholipid components did not indicate any bilayer perturbation, so the formation of toroidal wormholes or micellarization can be excluded as a mechanism of its antimicrobial or cell penetrating action. However, 2H-NMR analysis of the acyl chain order parameter profiles showed that BP100 leads to considerable membrane thinning and thereby local destabilization.

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Acknowledgments

We acknowledge A. Eisele and K. Scheubeck (KIT) for the technical support with peptide synthesis, R. A. M. Ciriello, C. Mink (KIT) for participation in the NMR measurements, P. K. Mykhailiuk and I. V. Komarov (University of Kyiv) for custom-synthesis of the amino acid CF3-Bpg. This research was supported by the DFG-Center for Functional Nanostructures (E1.2).

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Authors and Affiliations

  1. Institute of Organic Chemistry, Karlsruhe Institute of Technology (KIT), Fritz-Haber-Weg 6, 76131, Karlsruhe, Germany

    Julia Misiewicz & Anne S. Ulrich

  2. Institute of Biological Interfaces (IBG-2), Karlsruhe Institute of Technology (KIT), POB 3640, 76021, Karlsruhe, Germany

    Sergii Afonin, Stephan L. Grage, Jonas van den Berg, Erik Strandberg, Parvesh Wadhwani & Anne S. Ulrich

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  1. Julia Misiewicz
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  2. Sergii Afonin
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  3. Stephan L. Grage
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  4. Jonas van den Berg
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  6. Parvesh Wadhwani
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  7. Anne S. Ulrich
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Correspondence to Anne S. Ulrich.

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Misiewicz, J., Afonin, S., Grage, S.L. et al. Action of the multifunctional peptide BP100 on native biomembranes examined by solid-state NMR. J Biomol NMR 61, 287–298 (2015). https://doi.org/10.1007/s10858-015-9897-8

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