2D IR Line Shapes Probe Ovispirin Peptide Conformation and Depth in Lipid Bilayers

Title2D IR Line Shapes Probe Ovispirin Peptide Conformation and Depth in Lipid Bilayers
Publication TypeJournal Article
Year of Publication2010
AuthorsWoys AMarie, Lin Y-S, Reddy AS, Xiong W, De Pablo JJ, Skinner JL, Zanni MT
Secondary TitleJournal of the American Chemical Society
Volume132
Issue8
Pagination2832 - 2838
Date Published03/2010
Abstract

We report a structural study on the membrane binding of ovispirin using 2D IR line shape analysis, isotope labeling, and molecular dynamics simulations. Ovispirin is an antibiotic polypeptide that binds to the surfaces of membranes as an α-helix. By resolving individual backbone vibrational modes (amide I) using 1-13C═18O labeling, we measured the 2D IR line shapes for 15 of the 18 residues in this peptide. A comparison of the line shapes reveals an oscillation in the inhomogeneous line width that has a period equal to that of an α-helix (3.6 amino acids). The periodic trend is caused by the asymmetric environment of the membrane bilayer that exposes one face of the α-helix to much stronger environmental electrostatic forces than the other. We compare our experimental results to 2D IR line shapes calculated using the lowest free energy structure identified from molecular dynamics simulations. These simulations predict a periodic trend similar to the experiment and lead us to conclude that ovispirin lies in the membrane just below the headgroups, is tilted, and may be kinked. Besides providing insight into the antibiotic mechanism of ovispirin, our procedure provides an infrared method for studying peptide and protein structures that relies on the natural vibrational modes of the backbone. It is a complementary method to other techniques that utilize line shapes, such as fluorescence, NMR, and ESR spectroscopies, because it does not require mutations, the spectra can be quantitatively simulated using molecular dynamics, and the technique can be applied to difficult-to-study systems like ion channels, aggregated proteins, and kinetically evolving systems.

URLhttp://pubs.acs.org/doi/abs/10.1021/ja9101776
DOI10.1021/ja9101776
Short TitleJ. Am. Chem. Soc.