Phospholipase C-induced aggregation and fusion of cholesterol-lecithin small unilamellar vesicles

We have investigated the effects of the Ca2+ -requiring enzyme phospholipase C on the stability of sonicated vesicles made with different molar ratios of cholesterol to lecithin. Vesicle aggregation is detected by following turbidity with time. Upon the addition of phospholipase C and after a short...

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Veröffentlicht in:Biochemistry (Easton) 1993-07, Vol.32 (27), p.6965-6973
Hauptverfasser: Luk, Andrew S, Kaler, Eric W, Lee, Sum P
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creator Luk, Andrew S
Kaler, Eric W
Lee, Sum P
description We have investigated the effects of the Ca2+ -requiring enzyme phospholipase C on the stability of sonicated vesicles made with different molar ratios of cholesterol to lecithin. Vesicle aggregation is detected by following turbidity with time. Upon the addition of phospholipase C and after a short lag period, the turbidity of a vesicle dispersion increases continuously with time. The rate of increase of turbidity increases with both the enzyme-to-vesicle ratio and the cholesterol content of the vesicles. Vesicle fusion and leakage of contents are monitored by a contents-mixing fusion assay using 8-aminonaphthalene-1,3,6- trisulfonic acid (ANTS) and p-xylyienebis(pyridinium bromide) (DPX) as the fluorescence probes [Ellens, H., Bentz, J., and Szoka, F.C. (1985) Biochemistry 24, 3099-3106]. The results clearly show that phospholipase C induces vesicle fusion. The rate of vesicle fusion correlates with the enzyme-to-vesicle ratio but not with the cholesterol content of the membrane. Negligible aggregation and fusion of vesicles occurs when the experiment is repeated with buffer free of Ca2+. The membrane-destabilizing diacylglycerol, a product of lecithin hydrolysis by phospholipase C, is speculated to play a major role in driving the observed vesicle aggregation and fusion. The kinetics of vesicle aggregation and vesicle fusion can be predicted by linking Michaelis-Menten enzyme kinetics to a mass-action model
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Psychology</subject><subject>GLICERIDOS</subject><subject>GLYCERIDE</subject><subject>Kinetics</subject><subject>LECITHINE</subject><subject>LECITINAS</subject><subject>Lipid Bilayers</subject><subject>MEMBRANA</subject><subject>MEMBRANE</subject><subject>Membrane Fusion</subject><subject>Membrane physicochemistry</subject><subject>Molecular biophysics</subject><subject>Naphthalenes - chemistry</subject><subject>Phosphatidylcholines - metabolism</subject><subject>Pyridinium Compounds - chemistry</subject><subject>Type C Phospholipases - metabolism</subject><issn>0006-2960</issn><issn>1520-4995</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1993</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpt0M1r2zAYBnAxNrI022m3wcCH0R6KN33Zso8jtGkh0EDb0Zt4I8mJOtlKpbi0__1kHEIPPenj-fHy8iD0jeBfBFPye20xxqICTOkHNCUFxTmv6-Ijmqb_Mqd1iT-jkxgf05NjwSdoUjHGCS2nCFZbH3db7-wOosnmue10r4zOYLMJZgN767sMOp01fRyuvslU0ibuTfAud0bZ_dZ2WWzBuazvrIPWOAchezbRqgS_oE8NuGi-Hs4Zur-8uJtf5cubxfX8zzIHjuk-p0RhQfCaC8W00bjSQjec0aoQRdloIkBRzSgtwRhcASeDXCfCBS1rXbEZOh3n7oJ_6tOCsrVRDbt0xvdRiiKNKnGR4PkIVfAxBtPIXbAthFdJsBwKlW8KTfrHYWy_bo0-2kODKf95yCEqcE2ATtl4ZLzivCYDy0dmU3MvxxjCP1kKJgp5t7qVf-mSPxSLB8mS_z76BryETUgj729rzpjgQ3g2hqCifPR96FKx727_H9xEodU</recordid><startdate>19930713</startdate><enddate>19930713</enddate><creator>Luk, Andrew S</creator><creator>Kaler, Eric W</creator><creator>Lee, Sum P</creator><general>American Chemical Society</general><scope>FBQ</scope><scope>BSCLL</scope><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope></search><sort><creationdate>19930713</creationdate><title>Phospholipase C-induced aggregation and fusion of cholesterol-lecithin small unilamellar vesicles</title><author>Luk, Andrew S ; Kaler, Eric W ; Lee, Sum P</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-a402t-21c0710b47c3ded08d7df43285756fd17ac2d3226aee08a41b47cbdf447269d83</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1993</creationdate><topic>ACTIVIDAD ENZIMATICA</topic><topic>ACTIVITE ENZYMATIQUE</topic><topic>Artificial membranes and reconstituted systems</topic><topic>Biological and medical sciences</topic><topic>CHOLESTEROL</topic><topic>Cholesterol - metabolism</topic><topic>COLESTEROL</topic><topic>ESTERASAS</topic><topic>ESTERASE</topic><topic>Fluorescent Dyes</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>GLICERIDOS</topic><topic>GLYCERIDE</topic><topic>Kinetics</topic><topic>LECITHINE</topic><topic>LECITINAS</topic><topic>Lipid Bilayers</topic><topic>MEMBRANA</topic><topic>MEMBRANE</topic><topic>Membrane Fusion</topic><topic>Membrane physicochemistry</topic><topic>Molecular biophysics</topic><topic>Naphthalenes - chemistry</topic><topic>Phosphatidylcholines - metabolism</topic><topic>Pyridinium Compounds - chemistry</topic><topic>Type C Phospholipases - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Luk, Andrew S</creatorcontrib><creatorcontrib>Kaler, Eric W</creatorcontrib><creatorcontrib>Lee, Sum P</creatorcontrib><collection>AGRIS</collection><collection>Istex</collection><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Biochemistry (Easton)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Luk, Andrew S</au><au>Kaler, Eric W</au><au>Lee, Sum P</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Phospholipase C-induced aggregation and fusion of cholesterol-lecithin small unilamellar vesicles</atitle><jtitle>Biochemistry (Easton)</jtitle><addtitle>Biochemistry</addtitle><date>1993-07-13</date><risdate>1993</risdate><volume>32</volume><issue>27</issue><spage>6965</spage><epage>6973</epage><pages>6965-6973</pages><issn>0006-2960</issn><eissn>1520-4995</eissn><abstract>We have investigated the effects of the Ca2+ -requiring enzyme phospholipase C on the stability of sonicated vesicles made with different molar ratios of cholesterol to lecithin. 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The membrane-destabilizing diacylglycerol, a product of lecithin hydrolysis by phospholipase C, is speculated to play a major role in driving the observed vesicle aggregation and fusion. The kinetics of vesicle aggregation and vesicle fusion can be predicted by linking Michaelis-Menten enzyme kinetics to a mass-action model</abstract><cop>Washington, DC</cop><pub>American Chemical Society</pub><pmid>8334126</pmid><doi>10.1021/bi00078a022</doi><tpages>9</tpages></addata></record>
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subjects ACTIVIDAD ENZIMATICA
ACTIVITE ENZYMATIQUE
Artificial membranes and reconstituted systems
Biological and medical sciences
CHOLESTEROL
Cholesterol - metabolism
COLESTEROL
ESTERASAS
ESTERASE
Fluorescent Dyes
Fundamental and applied biological sciences. Psychology
GLICERIDOS
GLYCERIDE
Kinetics
LECITHINE
LECITINAS
Lipid Bilayers
MEMBRANA
MEMBRANE
Membrane Fusion
Membrane physicochemistry
Molecular biophysics
Naphthalenes - chemistry
Phosphatidylcholines - metabolism
Pyridinium Compounds - chemistry
Type C Phospholipases - metabolism
title Phospholipase C-induced aggregation and fusion of cholesterol-lecithin small unilamellar vesicles
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