Biophys J, August 2002, p. 1004-1013, Vol. 83, No. 2

Membrane Composition Determines Pardaxin's Mechanism of Lipid Bilayer Disruption

Kevin J. Hallock,^* Dong-Kuk Lee,^* John Omnaas,^§ Henry I. Mosberg,^§ and A. Ramamoorthy^*

 ^*Department of Chemistry,  Biophysics Research Division,  Macromolecular Science and Engineering,  ^§College of Pharmacy, University of Michigan, Ann Arbor, Michigan 48109 USA

Pardaxin is a membrane-lysing peptide originally isolated from the fish Pardachirus marmoratus. The effect of the carboxy-amide of pardaxin (P1a) on bilayers of varying composition was studied using ¹⁵N and ³¹P solid-state NMR of mechanically aligned samples and differential scanning calorimetry (DSC). ¹⁵N NMR spectroscopy of [¹⁵N-Leu₁₉]P1a found that the orientation of the peptide's C-terminal helix depends on membrane composition. It is located on the surface of lipid bilayers composed of 1-palmitoyl-2-oleoyl-phosphatidylcholine (POPC) and is inserted in lipid bilayers composed of 1,2-dimyristoyl-phosphatidylcholine (DMPC). The former suggests a carpet mechanism for bilayer disruption whereas the latter is consistent with a barrel-stave mechanism. The ³¹P chemical shift NMR spectra showed that the peptide significantly disrupts lipid bilayers composed solely of zwitterionic lipids, particularly bilayers composed of POPC, in agreement with a carpet mechanism. P1a caused the formation of an isotropic phase in 1-palmitoyl-2-oleoyl-phosphatidylethanolamine (POPE) lipid bilayers. This, combined with DSC data that found P1a reduced the fluid lamellar-to-inverted hexagonal phase transition temperature at very low concentrations (1:50,000), is interpreted as the formation of a cubic phase and not micellization of the membrane. Experiments exploring the effect of P1a on lipid bilayers composed of 4:1 POPC:cholesterol, 4:1 POPE:cholesterol, 3:1 POPC:1-palmitoyl-2-oleoyl-phosphatidylglycerol (POPG), and 3:1 POPE:POPG were also conducted, and the presence of anionic lipids or cholesterol was found to reduce the peptide's ability to disrupt bilayers. Considered together, these data demonstrate that the mechanism of P1a is dependent on membrane composition.

Biophys J, August 2002, p. 1004-1013, Vol. 83, No. 2
© 2002 by the Biophysical Society   0006-3495/02/08/1004/10  $2.00

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