Induction of Morphological Changes in Model Lipid Membranes and the Mechanism of Membrane Disruption by a Large Scorpion-Derived Pore-Forming Peptide (original) (raw)

2005, Biophysical Journal

The membrane disruption mechanism of pandinin 1 (pin1), an antimicrobial peptide isolated from the venom of the African scorpion, was studied using 31 P, 13 C, 1 H solid-state and multidimensional solution-state NMR spectroscopy. A highresolution NMR solution structure of pin1 showed that the two distinct a-helical regions move around the central hinge region, which contains Pro 19 . 31 P NMR spectra of lipid membrane in the presence of pin1, at various temperatures, showed that pin1 induces various lipid phase behaviors depending on the acyl chain length and charge of phospholipids. Notably, it was found that pin1 induced formation of the cubic phase in shorter lipid membranes above T m . Further, the 13 C NMR spectra of pin1 labeled at Leu 28 under magic angle spinning (MAS) indicated that the motion of pin1 bound to the lipid bilayer was very slow, with a correlation time of the order of 10 ÿ3 s. 31 P NMR spectra of dispersions of four saturated phosphatidyl-cholines in the presence of three types of pin1 derivatives, [W4A, W6A, W15A]-pin1, pin1(1-18), and pin1(20-44), at various temperatures demonstrated that all three pin1 derivatives have a reduced ability to trigger the cubic phase. 13 C chemical shift values for pin1(1-18) labeled at Val 3 , Ala 10 , or Ala 11 under static or slow MAS conditions indicate that pin1(1-18) rapidly rotates around the average helical axis, and the helical rods are inclined at ;30°to the lipid long axis. 13 C chemical shift values for pin1(20-44) labeled at Gly 25 , Leu 28 , or Ala 31 under static conditions indicate that pin1(20-44) may be isotropically tumbling. 1 H MAS chemical shift measurements suggest that pin1 is located at the membrane-water interface approximately parallel to the bilayer surface. Solid-state NMR results correlated well with the observed biological activity of pin1 in red blood cells and bacteria.