Monitoring phosphatidic acid formation in intact phosphatidylcholine bilayers upon phospholipase D catalysis.

ABSTRACT: We have monitored the production of the negatively charged lipid, 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphatidic acid acid (POPA), in supported lipid bilayers via the enzymatic hydrolysis of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (PC), a zwitterionic lipid. Experiments were performed with phospholipase D (PLD) in a Ca(2+) dependent fashion. The strategy for doing this involved using membrane-bound streptavidin as a biomarker for the charge on the membrane. The focusing position of streptavidin in electrophoretic-electroosmotic focusing (EEF) experiments was monitored via a fluorescent tag on this protein. The negative charge increased during these experiments due to the formation of POPA lipids. This caused the focusing position of streptavidin to migrate toward the negatively ch

SUBMITTER: Liu C 

PROVIDER: S-EPMC3983022 | biostudies-literature | 2014 Feb

REPOSITORIES: biostudies-literature