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- Scopus: eid_2-s2.0-0027407050
- PMID: 8429256
- WOS: WOS:A1993KL95900005
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Article: Lipid vesicle fusion induced by phospholipase C activity in model bile
Title | Lipid vesicle fusion induced by phospholipase C activity in model bile |
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Authors | |
Keywords | biliary vesicles diacylglycerol fluorescence spectroscopy gallstones liposomes membrane fusion quasielastic light scattering spectroscopy |
Issue Date | 1993 |
Publisher | American Society for Biochemistry and Molecular Biology, Inc. The Journal's web site is located at http://www.jlr.org/ |
Citation | Journal Of Lipid Research, 1993, v. 34 n. 2, p. 211-217 How to Cite? |
Abstract | Using a system of phosphatidylcholine-cholesterol vesicles to model the vesicle phase of mammalian bile (1:1 molar ratio) we evaluated whether very small amounts of C. perfringens phospholipase C activity (0.5-6.5 nmol/min per ml) could lead to vesicle fusion, a precursor step for cholesterol precipitation in gallbladder bile. Quasielastic light scattering spectroscopy (QLS) was used to monitor vesicle growth and aggregation in model bile (0.89 mM total lipid) in the presence of phospholipase C. Vesicle growth over 2 h could be detected with phospholipase activity as little as 0.5 nmol/min per ml. Vesicle growth was sustainable over days in the absence of Ca2+ once as little as 3-7 mol% diacylglycerol had been generated as a result of the initial phospholipase C treatment. The presence of fusion intermediates was confirmed using transmission electron microscopy. In addition, kinetically slow vesicle fusion with intravesicle content mixing and minimal leakage was also confirmed by fluorescence spectroscopy using two populations of vesicles containing 5 mM TbCl3 or 50 mM dipicolinic acid. Efficient fusion (40% maximum fluorescence) was obtained at 30 min at 25°C with phospholipase C activity. This level of enzyme activity approximates that found in human gallbladder bile (1.2 nmol/min per ml). We conclude that the hydrolysis products of phospholipase C activity can, in very small amounts (3-7 mol% diacylglycerol), lead to destabilization and fusion of cholesterol-saturated biliary vesicles. A reappraisal of the importance of phospholipase C hydrolysis products in the pathogenesis of cholesterol gallstones is warranted based on these observations. |
Persistent Identifier | http://hdl.handle.net/10722/175684 |
ISSN | 2023 Impact Factor: 5.0 2023 SCImago Journal Rankings: 2.090 |
ISI Accession Number ID |
DC Field | Value | Language |
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dc.contributor.author | Little, TE | en_US |
dc.contributor.author | Madani, H | en_US |
dc.contributor.author | Lee, SP | en_US |
dc.contributor.author | Kaler, EW | en_US |
dc.date.accessioned | 2012-11-26T09:00:29Z | - |
dc.date.available | 2012-11-26T09:00:29Z | - |
dc.date.issued | 1993 | en_US |
dc.identifier.citation | Journal Of Lipid Research, 1993, v. 34 n. 2, p. 211-217 | en_US |
dc.identifier.issn | 0022-2275 | en_US |
dc.identifier.uri | http://hdl.handle.net/10722/175684 | - |
dc.description.abstract | Using a system of phosphatidylcholine-cholesterol vesicles to model the vesicle phase of mammalian bile (1:1 molar ratio) we evaluated whether very small amounts of C. perfringens phospholipase C activity (0.5-6.5 nmol/min per ml) could lead to vesicle fusion, a precursor step for cholesterol precipitation in gallbladder bile. Quasielastic light scattering spectroscopy (QLS) was used to monitor vesicle growth and aggregation in model bile (0.89 mM total lipid) in the presence of phospholipase C. Vesicle growth over 2 h could be detected with phospholipase activity as little as 0.5 nmol/min per ml. Vesicle growth was sustainable over days in the absence of Ca2+ once as little as 3-7 mol% diacylglycerol had been generated as a result of the initial phospholipase C treatment. The presence of fusion intermediates was confirmed using transmission electron microscopy. In addition, kinetically slow vesicle fusion with intravesicle content mixing and minimal leakage was also confirmed by fluorescence spectroscopy using two populations of vesicles containing 5 mM TbCl3 or 50 mM dipicolinic acid. Efficient fusion (40% maximum fluorescence) was obtained at 30 min at 25°C with phospholipase C activity. This level of enzyme activity approximates that found in human gallbladder bile (1.2 nmol/min per ml). We conclude that the hydrolysis products of phospholipase C activity can, in very small amounts (3-7 mol% diacylglycerol), lead to destabilization and fusion of cholesterol-saturated biliary vesicles. A reappraisal of the importance of phospholipase C hydrolysis products in the pathogenesis of cholesterol gallstones is warranted based on these observations. | en_US |
dc.language | eng | en_US |
dc.publisher | American Society for Biochemistry and Molecular Biology, Inc. The Journal's web site is located at http://www.jlr.org/ | en_US |
dc.relation.ispartof | Journal of Lipid Research | en_US |
dc.subject | biliary vesicles | - |
dc.subject | diacylglycerol | - |
dc.subject | fluorescence spectroscopy | - |
dc.subject | gallstones | - |
dc.subject | liposomes | - |
dc.subject | membrane fusion | - |
dc.subject | quasielastic light scattering spectroscopy | - |
dc.subject.mesh | Bile - Chemistry | en_US |
dc.subject.mesh | Calcium - Pharmacology | en_US |
dc.subject.mesh | Clostridium Perfringens - Enzymology | en_US |
dc.subject.mesh | Crystallization | en_US |
dc.subject.mesh | Edetic Acid - Pharmacology | en_US |
dc.subject.mesh | Light | en_US |
dc.subject.mesh | Lipid Metabolism | en_US |
dc.subject.mesh | Liposomes - Metabolism | en_US |
dc.subject.mesh | Membrane Fusion | en_US |
dc.subject.mesh | Microscopy, Electron | en_US |
dc.subject.mesh | Particle Size | en_US |
dc.subject.mesh | Phosphatidylcholines - Metabolism | en_US |
dc.subject.mesh | Scattering, Radiation | en_US |
dc.subject.mesh | Spectrometry, Fluorescence | en_US |
dc.subject.mesh | Spectrum Analysis | en_US |
dc.subject.mesh | Type C Phospholipases - Metabolism | en_US |
dc.title | Lipid vesicle fusion induced by phospholipase C activity in model bile | en_US |
dc.type | Article | en_US |
dc.identifier.email | Lee, SP: sumlee@hku.hk | en_US |
dc.identifier.authority | Lee, SP=rp01351 | en_US |
dc.description.nature | link_to_subscribed_fulltext | en_US |
dc.identifier.pmid | 8429256 | - |
dc.identifier.scopus | eid_2-s2.0-0027407050 | en_US |
dc.identifier.volume | 34 | en_US |
dc.identifier.issue | 2 | en_US |
dc.identifier.spage | 211 | en_US |
dc.identifier.epage | 217 | en_US |
dc.identifier.isi | WOS:A1993KL95900005 | - |
dc.publisher.place | United States | en_US |
dc.identifier.scopusauthorid | Little, TE=35822432000 | en_US |
dc.identifier.scopusauthorid | Madani, H=6701613342 | en_US |
dc.identifier.scopusauthorid | Lee, SP=7601417497 | en_US |
dc.identifier.scopusauthorid | Kaler, EW=7007157989 | en_US |
dc.identifier.issnl | 0022-2275 | - |