This Article Full Text Full Text (PDF) All Versions of this Article: biophysj.105.070292v1 89/6/4067    most recent Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via Google Scholar Google Scholar Articles by Nomura, K. Articles by Corzo, G. Search for Related Content PubMed PubMed Citation Articles by Nomura, K. Articles by Corzo, G.

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

Kaoru Nomura ^*, Gilles Ferrat ^* , Terumi Nakajima ^*, Herve Darbon , Takashi Iwashita ^* and Gerardo Corzo ^* 

^* Suntory Institute for Bioorganic Research, Osaka 618-8503, Japan; AFMB, CNRS UMR 6098 and Universités d'Aix-Marseille I and II, 13402 Marseille Cedex 20, France; and Institute of Biotechnology-UNAM, Cuernavaca, Morelos 62210, Mexico

Correspondence: Address reprint requests to K. Nomura, E-mail: nomura{at}sunbor.or.jp; or to G. Corzo, E-mail: corzo{at}ibt.unam.mx.

The membrane disruption mechanism of pandinin 1 (pin1), an antimicrobial peptide isolated from the venom of the African scorpion, was studied using ³¹P, ¹³C, ¹H solid-state and multidimensional solution-state NMR spectroscopy. A high-resolution NMR solution structure of pin1 showed that the two distinct -helical regions move around the central hinge region, which contains Pro¹⁹. ³¹P 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 ¹³C NMR spectra of pin1 labeled at Leu²⁸ 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^–3 s. ³¹P 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. ¹³C chemical shift values for pin1(1-18) labeled at Val³, Ala¹⁰, or Ala¹¹ 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. ¹³C chemical shift values for pin1(20-44) labeled at Gly²⁵, Leu²⁸, or Ala³¹ under static conditions indicate that pin1(20-44) may be isotropically tumbling. ¹H 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.