In this work, we report the design, synthesis, and application of a bis-pyrene phospholipid probe for detection of phospholipase A action through changes in pyrene monomer and excimer fluorescence intensities. Continuous fluorometric assays enabled detection of the activities of multiple PLA enzymes as well as the decrease in catalysis by PLA from honey bee venom caused by the inhibitor p-bromo phenacylbromide. Thin-layer chromatography and mass spectrometry analysis were also used to validate probe hydrolysis by PLA. Mass spectrometry data also supported cleavage of the probe by phospholipase C and D enzymes, although changes in fluorescence were not observed in these cases. Nevertheless, the bis-pyrene phospholipid probe developed in this work is effective for detection of PLA enzyme activity through an assay that enables screening for inhibitor development.
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Development of a bis-pyrene phospholipid probe for fluorometric detection of phospholipase A inhibition.
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Department of Chemistry, University of Tennessee, 1420 Circle Drive, Knoxville, TN 37996 USA. Electronic address:
In this work, we report the design, synthesis, and application of a bis-pyrene phospholipid probe for detection of phospholipase A action through changes in pyrene monomer and excimer fluorescence intensities. Continuous fluorometric assays enabled detection of the activities of multiple PLA enzymes as well as the decrease in catalysis by PLA from honey bee venom caused by the inhibitor p-bromo phenacylbromide. Thin-layer chromatography and mass spectrometry analysis were also used to validate probe hydrolysis by PLA.
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School of Chemistry , University of Bristol, Cantock's Close , Bristol BS8 1TS , UK . Email:
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