Investigation into the role of phosphatidylserine in modifying the susceptibility of human lymphocytes to secretory phospholipase A₂ using cells deficient in the expression of scramblase

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• 作者：Jennifer Nelson ; Lyndee L. Francom ; Lynn Anderson ; Kelly Damm ; Ryan Baker ; Joseph Chen ; Sarah Franklin ; Amy Hamaker ; Izadora Izidoro ; Eric Moss ; Mikayla Orton ; Evan Stevens ; Celestine Yeung ; Allan M. Judd ; John D. Bell ; ^{john_bell@byu.edu}
• 关键词：Secretory phospholipase A2 ; Phosphatidylserine exposure ; Biophysical membrane change ; Hydrolysis ; Merocyanine 540 ; Trimethylammonium diphenylhexatriene
• 刊名：Biochimica et Biophysica Acta (BBA)/Biomembranes
• 出版年：2012
• 出版时间：May, 2012
• 年：2012
• 卷：1818
• 期：5
• 页码：1196-1204
• 全文大小：915 K

文摘

Normal human lymphocytes resisted the hydrolytic action of secretory phospholipase A₂ but became susceptible to the enzyme following treatment with a calcium ionophore, ionomycin. To test the hypothesis that this susceptibility requires exposure of the anionic lipid phosphatidylserine on the external face of the cell membrane, experiments were repeated with a human Burkitt's lymphoma cell line (Raji cells). In contrast to normal lymphocytes or S49 mouse lymphoma cells, most of the Raji cells (83 % ) did not translocate phosphatidylserine to the cell surface upon treatment with ionomycin. Those few that did display exposed phosphatidylserine were hydrolyzed immediately upon addition of phospholipase A₂. Interestingly, the remaining cells were also completely susceptible to the enzyme but were hydrolyzed at a slower rate and after a latency of about 100 s. In contradistinction to the defect in phosphatidylserine translocation, Raji cells did display other physical membrane changes upon ionomycin treatment that may be relevant to hydrolysis by phospholipase A₂. These changes were detected by merocyanine 540 and trimethylammonium diphenylhexatriene fluorescence and were common among normal lymphocytes, S49 cells, and Raji cells. The levels of these latter effects corresponded well with the relative rates of hydrolysis among the three cell lines. These results suggested that while phosphatidylserine enhances the rate of cell membrane hydrolysis by secretory phospholipase A₂, it is not an absolute requirement. Other physical properties such as membrane order contribute to the level of membrane susceptibility to the enzyme independent of phosphatidylserine.