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Investigating the effect of a single glycine to alanine substitution on interactions of antimicrobial peptide latarcin 2a with a lipid membrane

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Abstract

Latarcins are linear, α-helical antimicrobial peptides purified from the venom of the Central Asian spider Lachesana tarabaevi, with lytic activity against Gram-positive and Gram-negative bacteria, erythrocytes, and yeast at micromolar concentrations. In this work, we investigated the role of the hinge in latarcin 2a (ltc2a, GLFGKLIKKFGRKAISYAVKKARGKH-COOH), which adopts a helix–hinge–helix conformation in membrane-mimicking environments, on peptide–membrane interactions and its potential effect on the selective toxicity of the peptide. A modified latarcin 2a, ltc2aG11A, obtained by replacing the glycine at position 11 with alanine (ltc2aG11A, GLFGKLIKKFARKAISYAVKKARGKH-COOH), adopts a more rigid structure due to the reduced conformational flexibility. Langmuir monolayer measurements combined with atomic force microscopy and X-ray photoemission electron microscopy (X-PEEM) indicate that both peptides bind and insert preferentially into anionic compared with zwitterionic phospholipid monolayers. Modified ltc2aG11A was found to be more disruptive of supported phospholipid bilayer modeling mammalian cell membrane. However, no considerable difference in lytic activity of the two peptides toward bacterial membrane was found. Overall the data indicate that decrease in the flexibility of ltc2a induced by the modification in the hinge region is likely to increase the peptide’s nonspecific interactions with zwitterionic cell membranes and potentially increase its toxicity against eukaryotic cells.

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Acknowledgments

Financial support was provided by ERA, NSERC, and CFI. X-ray microscopy was carried out using the polymer STXM on beamline 5.3.2 and the magnetic X-PEEM on beamline 7.3.1 at the ALS, which is supported by the US DoE under contract DE-AC03-76SF00098. We thank David Kilcoyne and Tolek Tyliszczak (STXM532) as well as Andreas Scholl and Andrew Doran (PEEM2) for their diligence and expertise in keeping the beamlines in top condition.

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Authors and Affiliations

  1. Department of Chemistry, Carleton University, 1125 Colonel By Dr., Ottawa, ON, K1S 5B6, Canada

    Grace Idiong, Amy Won, Annamaria Ruscito & Anatoli Ianoul

  2. Brockhouse Institute for Materials Research, McMaster University, Hamilton, ON, L8S 4M1, Canada

    Bonnie O. Leung & Adam P. Hitchcock

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  1. Grace Idiong
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  2. Amy Won
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  3. Annamaria Ruscito
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Correspondence to Anatoli Ianoul.

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Idiong, G., Won, A., Ruscito, A. et al. Investigating the effect of a single glycine to alanine substitution on interactions of antimicrobial peptide latarcin 2a with a lipid membrane. Eur Biophys J 40, 1087–1100 (2011). https://doi.org/10.1007/s00249-011-0726-z

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  • DOI: https://doi.org/10.1007/s00249-011-0726-z

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