Biophysical Journal 57: 351-362 (1990)
© 1990 the Biophysical Society

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Optimizing and characterizing alignment of oriented lipid bilayers containing gramicidin D.

Department of Chemistry, Florida State University, Tallahassee 32306-3006.

ABSTRACT

31P NMR spectroscopy and optical microscopy have been used to characterize samples of gramicidin D in oriented lipid bilayers. Correlations have been made between the defect structures observed under crossed polarizers by optical microscopy and characteristic features of 31P NMR spectra. The sample preparation protocol has been improved using these techniques to achieve minimal dispersion of the bilayer normal and minimal amounts of unoriented sample. The molar ratio of gramicidin to dimyristoyl-phosphatidylcholine, the extent of hydration, and the cosolubilizing solvent system were used as the protocol variables. While hydration level and solvent system had profound effects on the sample orientation the molar ratio did not. However, the 31P chemical shift anisotropy is very sensitive to the molar ratio and can be used as an in situ method for determining the molar ratio.

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