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Article: A fluorescence energy transfer study of lecithin-cholesterol vesicles in the presence of phospholipase C

TitleA fluorescence energy transfer study of lecithin-cholesterol vesicles in the presence of phospholipase C
Authors
KeywordsDansyl
Dehydroergosterol
Gallstones
Nucleation
Issue Date1999
PublisherAmerican Society for Biochemistry and Molecular Biology, Inc. The Journal's web site is located at http://www.jlr.org/
Citation
Journal Of Lipid Research, 1999, v. 40 n. 8, p. 1483-1494 How to Cite?
AbstractWe demonstrate Forster resonance energy transfer from dehydroergosterol to dansylated lecithin in lecithin-cholesterol vesicles and characterize the vesicles in the presence of the pro-nucleating enzyme, phospholipase C (PLC). Exposure to phospholipase C causes a temporary decrease in the dehydroergosterol to dansyl fluorescence ratio followed by an increase to and above the initial value. The temporary decrease in the fluorescence ratio results from an increase in the dansylated lecithin intensity that coincides with a dansyl blue shift. The extent of the blue shift correlates with the level of diacylglycerol generated in situ by PLC, suggesting an increased association between dansylated lecithin and cholesterol as membrane fluidity increases and membrane polarity decreases. The subsequent increase in the fluorescence ratio results from both an increase in the dehydroergsterol intensity and a concomitant decrease in the dansylated lecithin intensity of equal magnitude. This signifies a reduction in energy transfer from dehydroergosterol to dansylated lecithin and indicates an increased separation between the two fluorophores. The increase in the fluorescence ratio persists beyond the time scales for vesicle aggregation and fusion, as measured by turbidity, and precedes the onset of macroscopic cholesterol crystals observed with an optical microscope. Thus, the increased separation between dehydroergosterol and dansylated lecithin is consistent with a mechanism of cholesterol nucleation from the vesicles. Moreover, the onset and rate of increase in the fluorescence ratio correlate with the cholesterol:lecithin mole ratio of the vesicles. Fluorescence energy transfer from dehydroergosterol to dansylated lecithin therefore shows potential as a methodology for measuring cholesterol nucleation in model bile.
Persistent Identifierhttp://hdl.handle.net/10722/92486
ISSN
2015 Impact Factor: 4.368
2015 SCImago Journal Rankings: 2.529
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorWrenn, SPen_HK
dc.contributor.authorKaler, EWen_HK
dc.contributor.authorLee, SPen_HK
dc.date.accessioned2010-09-17T10:47:43Z-
dc.date.available2010-09-17T10:47:43Z-
dc.date.issued1999en_HK
dc.identifier.citationJournal Of Lipid Research, 1999, v. 40 n. 8, p. 1483-1494en_HK
dc.identifier.issn0022-2275en_HK
dc.identifier.urihttp://hdl.handle.net/10722/92486-
dc.description.abstractWe demonstrate Forster resonance energy transfer from dehydroergosterol to dansylated lecithin in lecithin-cholesterol vesicles and characterize the vesicles in the presence of the pro-nucleating enzyme, phospholipase C (PLC). Exposure to phospholipase C causes a temporary decrease in the dehydroergosterol to dansyl fluorescence ratio followed by an increase to and above the initial value. The temporary decrease in the fluorescence ratio results from an increase in the dansylated lecithin intensity that coincides with a dansyl blue shift. The extent of the blue shift correlates with the level of diacylglycerol generated in situ by PLC, suggesting an increased association between dansylated lecithin and cholesterol as membrane fluidity increases and membrane polarity decreases. The subsequent increase in the fluorescence ratio results from both an increase in the dehydroergsterol intensity and a concomitant decrease in the dansylated lecithin intensity of equal magnitude. This signifies a reduction in energy transfer from dehydroergosterol to dansylated lecithin and indicates an increased separation between the two fluorophores. The increase in the fluorescence ratio persists beyond the time scales for vesicle aggregation and fusion, as measured by turbidity, and precedes the onset of macroscopic cholesterol crystals observed with an optical microscope. Thus, the increased separation between dehydroergosterol and dansylated lecithin is consistent with a mechanism of cholesterol nucleation from the vesicles. Moreover, the onset and rate of increase in the fluorescence ratio correlate with the cholesterol:lecithin mole ratio of the vesicles. Fluorescence energy transfer from dehydroergosterol to dansylated lecithin therefore shows potential as a methodology for measuring cholesterol nucleation in model bile.en_HK
dc.languageengen_HK
dc.publisherAmerican Society for Biochemistry and Molecular Biology, Inc. The Journal's web site is located at http://www.jlr.org/en_HK
dc.relation.ispartofJournal of Lipid Researchen_HK
dc.subjectDansylen_HK
dc.subjectDehydroergosterolen_HK
dc.subjectGallstonesen_HK
dc.subjectNucleationen_HK
dc.titleA fluorescence energy transfer study of lecithin-cholesterol vesicles in the presence of phospholipase Cen_HK
dc.typeArticleen_HK
dc.identifier.emailLee, SP: sumlee@hku.hken_HK
dc.identifier.authorityLee, SP=rp01351en_HK
dc.description.naturelink_to_subscribed_fulltext-
dc.identifier.pmid10428985-
dc.identifier.scopuseid_2-s2.0-0032776578en_HK
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-0032776578&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume40en_HK
dc.identifier.issue8en_HK
dc.identifier.spage1483en_HK
dc.identifier.epage1494en_HK
dc.identifier.isiWOS:000081865200013-
dc.publisher.placeUnited Statesen_HK
dc.identifier.scopusauthoridWrenn, SP=6603940041en_HK
dc.identifier.scopusauthoridKaler, EW=7007157989en_HK
dc.identifier.scopusauthoridLee, SP=7601417497en_HK

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