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Location of two antioxidants in oriented model membranes. Small-angle x-ray diffraction study.

Biophysical Journal

X-Ray Diffraction, chemistry, Vitamin E, Lipid Bilayers, Fourier Analysis, Cholesterol, Butylated Hydroxytoluene, Biophysics, Biophysical Phenomena, Antioxidants, 1,2-Dipalmitoylphosphatidylcholine

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      Small-angle x-ray diffraction has been applied in locating either butylated hydroxytoluene (BHT) or delta-tocopherol and their brominated analogues at a concentration of 40 mol% in oriented bilayers of dipalmitoylphosphatidylcholine (DPPC) or DPPC + 15 mol% cholesterol at 20 degrees C. Phases were determined using swelling experiments with structure factors plotted in reciprocal space, creating a relatively smooth curve as the amount of water between the bilayers was changed. Continuous Fourier transforms were also calculated using sampling theory (Shannon, C. E. 1949. Proc. Inst. Radio Engrs. NY. 37:10-21) to further test the consistency of the phase assignments. Fourier synthesis of structure factors resulted in absolute electron density profiles for different bilayers to a resolution of 5-6 A. In addition, difference Patterson maps were constructed to confirm the positions of the bromine atoms in the unit cell. Analysis of the data indicates the following: (a) The BHT molecules are dispersed throughout the alkyl-chain region in DPPC samples with and without cholesterol. (b) The chromanol ring of delta-tocopherol is in the vicinity of the glycerol backbone-headgroup region in samples of DPPC or DPPC + 15 mol% cholesterol. (c) Difference Patterson maps confirm the localization of bromine atoms in the various delta-tocopherol samples and lack of bromine localization in the various BHT samples.

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