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Biophys J. 2008 Apr 15;94(8):3084-93. doi: 10.1529/biophysj.107.118356. Epub 2008 Jan 11.

Effects of cholesterol on physical properties of human erythrocyte membranes: impact on susceptibility to hydrolysis by secretory phospholipase A2.

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Department of Physiology and Developmental Biology, Brigham Young University, Provo, Utah 84602, USA.


The ability of secretory phospholipase A(2) (sPLA(2)) to hydrolyze cell membranes is highly dependent on the physical properties of the membrane. The effects of cholesterol on these properties have been characterized in artificial bilayers and found to alter sPLA(2) activity significantly. It is hypothesized that the natural difference in cholesterol content between erythrocytes and leukocytes is in part responsible for their differing susceptibility to hydrolysis by sPLA(2). To test this hypothesis, defined amounts of cholesterol were removed from erythrocyte membranes using methyl-beta-cyclodextrin. Treatment of cells with methyl-beta-cyclodextrin increased the hydrolysis rate and total substrate hydrolyzed by sPLA(2). In general, this effect of cholesterol removal was more pronounced at higher temperatures. Comparison of the level of membrane order (assessed with the fluorescent probe laurdan) with hydrolysis rate revealed that sPLA(2) activity was greatly enhanced upon significant reductions in lipid order. Additional treatment of the cells with calcium ionophore further enhanced the hydrolysis rate and altered the relationship with membrane order. These data demonstrated that interactions with sPLA(2) observed in artificial bilayers apply to biological membranes. It is also proposed that the high level of cholesterol in erythrocyte membranes is a protective mechanism to guard against hydrolytic enzymes.

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