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X-ray scattering studies of model lipid membrane interacting with purothionin provide support for a previously proposed mechanism of membrane lysis

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Abstract

Thionins, ubiquitous plant toxins, are believed to act by lysing the membrane of pathogenic organisms. Several competing mechanisms were proposed for the lysis of phospholipid membranes by the toxins. In order to study in more detail the proposed mechanisms and possibly resolve among the competing proposals, the interactions of purothionins with a model lipid membrane in the form of a monolayer were studied. The monolayer formed at the air-water interface was studied by synchrotron X-ray reflectivity and grazing incidents diffraction methods. The model membrane was composed of 90:10 mol% DPPC:DPPS (dipylmitoyl phosphatidylcholine:dipylmitoyl phosphatidylserine). The protein interaction with the monolayer disturbs the in-plane and out-of-plane order of phospholipids, increases the amount of the liquid phase of the monolayer, and increases the average surface area per alkyl chain. The results indicate that the protein is bound only transiently, and after ~4 h most of the properties of the monolayer are reminiscent of the pure DPPC monolayer suggesting partial withdrawal of DPPS. Obtained electron density distributions perpendicular to the membrane interface do not show any significant contribution from the adsorbed proteins, further supporting the withdrawal hypothesis.

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Abbreviations

GIXD:

Grazing incidence X-ray diffraction

XR:

X-ray reflectivity

DPPC:

Dipylmitoyl phosphatidylcholine

DPPS:

Dipylmitoyl phosphatidylserine

PT:

Purothionin

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Acknowledgments

The work was supported by the Los Alamos National Laboratory under the auspices of the United States Department of Energy under DOE contract W7405-ENG-36, and by the DOE Office of Basic Energy Science. We thank Dr. Kristian Kjaer from the University of Copenhagen in Denmark who collaborated with us on the GIXD and XR measurements at HASYLAB, Hamburg, Germany.

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Authors and Affiliations

  1. Manuel Lujan Jr. Neutron Scattering Center, Los Alamos National Laboratory, MS-H805, Los Alamos, NM, 87545, USA

    Jaroslaw Majewski

  2. Infectious and Inflammatory Disease Center, Burnham Institute for Medical Research, 10901 N. Torrey Pines Rd., La Jolla, CA, 92037, USA

    Boguslaw Stec

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  1. Jaroslaw Majewski
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  2. Boguslaw Stec
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Correspondence to Boguslaw Stec.

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Majewski, J., Stec, B. X-ray scattering studies of model lipid membrane interacting with purothionin provide support for a previously proposed mechanism of membrane lysis. Eur Biophys J 39, 1155–1165 (2010). https://doi.org/10.1007/s00249-009-0568-0

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  • DOI: https://doi.org/10.1007/s00249-009-0568-0

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