Role of membrane oxidation in controlling the activity of human group IIa secretory phospholipase A₂ toward apoptotic lymphoma cells

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• 作者：Elizabeth Gibbons ; Jennifer Nelson ; Lynn Anderson ; Kelly Brewer ; Stephanie Melchor ; Allan M. Judd ; John D. Bell ; ^{john_bell@byu.edu}
• 关键词：Phospholipase A2 specificity ; Merocyanine 540 ; Hydrolysis kinetics ; Patman ; Trimethylammonium diphenylhexatriene ; C11-BODIPY
• 刊名：Biochimica et Biophysica Acta (BBA)/Biomembranes
• 出版年：2013
• 出版时间：February, 2013
• 年：2013
• 卷：1828
• 期：2
• 页码：670-676
• 全文大小：853 K

文摘

The membranes of healthy lymphocytes normally resist hydrolysis by secretory phospholipase A₂. However, they become susceptible during the process of apoptosis. Previous experiments have demonstrated the importance of certain physical changes to the membrane during cell death such as a reduction in membrane lipid order and exposure of phosphatidylserine on the membrane surface. Nevertheless, those investigations also showed that at least one additional factor was required for rapid hydrolysis by the human group IIa phospholipase isozyme. This study was designed to test the possibility that oxidation of membrane lipids is the additional factor. Flow cytometry and confocal microscopy with a fluorescent probe of oxidative potential suggested that oxidation of the plasma membrane occurs during apoptosis stimulated by thapsigargin. When oxidative potential was high, the activity of human group IIa secretory phospholipase A₂ was enhanced 30- to 100-fold compared to that observed with conditions sufficient for maximal hydrolysis by other secretory phospholipase A₂ isoforms. Direct oxidation of cell membranes with either of two oxidizing agents also stimulated hydrolysis by secretory phospholipase A₂. Both oxidizers caused externalization of phosphatidylserine, but a change in lipid order did not always occur. These results demonstrated that membrane oxidation strongly stimulates human group IIa secretory phospholipase A₂ activity toward apoptotic cells. Interestingly, the change in membrane order, previously thought to be imperative for high rates of hydrolysis, was not required when membrane lipids were oxidized. Whether phosphatidylserine exposure is still necessary with oxidation remains unresolved since the two events could not be deconvoluted.