Browsing by Subject "phospholipids"

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    AFFINITY OF CHOLESTEROL FOR POLYUNSATURATED FATTY ACID-CONTAINING PHOSPHOLIPIDS
    (Office of the Vice Chancellor for Research, 2012-04-13) Kagimbi, Maureen W.; Williams, Justin A.; Wassall, Stephen R.
    A wide range of health benefits is associated with the consumption of omega-3 polyunsaturated fatty acids (PUFAs). One possible mechanism is that through our diet, they are incorporated into the phospholipids of the plasma membrane and disrupt the molecular organization of membrane do-mains due to the high disorder of PUFA. Our focus is the interaction of PUFA with cholesterol, a major component in plasma membranes. The objective here is to measure the affinity of cholesterol for PUFA-containing phospholip-ids by observing how cholesterol partitions between large unilamellar vesi-cles (LUVs) and Cyclodextrin (CD). Crucial to this determination, we need to be able to determine the concentration of cholesterol in LUVs and CD using an enzymatic colorimetric assay to create a standard curve of light absorb-ance (at 570nm wavelength) as a function of cholesterol concentration. The assay and its application to measuring binding coefficients for cholesterol will be described.
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    An Investigation of Whether Vitamin E Preferentially Interacts with Polyunsaturated Lipids
    (Office of the Vice Chancellor for Research, 2015-04-17) Cavazos, Andres; Kinnun, Jacob J.; Williams, Justin A.; Bank, Morris; Ray, Bruce D.; Harper, Paul E.
    Vitamin E (α-tocopherol) is a lipid-soluble antioxidant that has the role of protecting phospholipids from oxidation in membranes. A question that remains is how the low concentration of α-tocopherol found in whole cells can protect the relatively large concentration of polyunsaturated phospholipids found in membranes that are particularly vulnerable to oxidative attack. We hypothesize that α-tocopherol colocalizes with polyunsaturated phospholipids to optimize its role as an antioxidant. This project attempts to test this hypothesis by comparing the effect of α-tocopherol on the molecular organization of 1-palmitoyl-2-docosahexaenoyl-sn-glycerophosphatidylethanolamine (16:0-22:6PE, PDPE) and, as a monounsaturated control, 1-palmitoyl-2-oleoyl-sn-glycerophosphatidylethanolamine (16:0-18:1PE, POPE) in mixtures with sphingomyelin (SM). By solid-state 2H NMR spectroscopy, we directly observe order and phase behavior of POPE-d31 and PDPE-d31 (analogs of POPE and PDPE with a perdeuterated sn-1 chain) in the mixed membranes. In complementary X-ray diffraction and differential scanning calorimetry experiments we further probe phase behavior. The spectra observed for POPE-d31 in POPE/SM/α-tocopherol (2:2:1 mol) reveal that a transition from gel to liquid crystalline phase is no longer apparent. At higher temperatures there is a superposition of two spectral components that we ascribe to α-tocopherol promoting a transition from lamellar to inverted hexagonal (HII) phase. Analysis of depaked spectra shows that order is increased by about 8 % and that the amount of HII phase increases with temperature, ranging from 7 (31 °C) to 41 % (65 °C). In mixed membranes where POPE-d31 is replaced by PDPE-d31, we shall investigate whether there is a greater tendency for α-tocopherol to increase order and destabilize bilayer structure for the polyunsaturated phospholipid.