Phospholipase A2 activity towards vesicles of DPPC and DMPC-DSPC containing small amounts of SMPC

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Phospholipase A2 (PLA2) is an interfacially active enzyme whose hydrolytic activity is known to be enhanced in one-component phospholipid bilayer substrates exhibiting dynamic micro-heterogeneity. In this study the activity of PLA2 towards large unilamellar vesicles composed of DPPC:SMPC and DMPC:DSPC:SMPC is investigated using fluorescence and HPLC techniques. Phase diagrams of the mixtures are established by differential scanning calorimetry and the PLA2 activity, monitored by the lag time, is correlated with the phase behavior of the mixtures. In addition, the degree of lipid hydrolysis in the DMPC:DSPC:SMPC lipid mixtures is detected by HPLC. The PLA2 activity is found to be significantly increased in the temperature range of the coexistence region where the lipid mixtures exhibit lateral gel-fluid phase separation. Furthermore, in the entire temperature range it is demonstrated that PLA2 preferentially hydrolyzes the short chain DMPC lipid. This discriminative effect becomes less pronounced when the asymmetric lipid SMPC is present in the lipid substrate. Inclusion of SMPC into either DPPC or DMPC:DSPC vesicles prolongs the lag time. The results clearly show that the PLA2 activity is significantly enhanced by lipid bilayer micro-heterogeneity in both one-component and multi-component lipid bilayer substrates. The PLA2 activity measurements are discussed in terms of dynamic gel-fluid lipid domain formation due to density fluctuations and static lipid domain formation due to gel-fluid phase separation.

Original languageEnglish
JournalBiochimica et Biophysica Acta - Biomembranes
Issue number2
Pages (from-to)133-143
Number of pages11
Publication statusPublished - 1 Dec 2001

    Research areas

  • Fluorescence, High performance liquid chromatography, Interfaces, Lipid bilayer, Micro-heterogeneity, Phase coexistence, Phase separation, Phospholipase A, Phospholipid

ID: 230987991