pH Alters PEG-mediated fusion of phosphatidylethanolamine-containing vesicles
Copyright © 2014 Biophysical Society. Published by Elsevier Inc. All rights reserved..
Here, we examine the different mechanisms of poly(ethylene glycol)-mediated fusion of small unilamellar vesicles composed of dioleoylphosphatidylcholine/dioleoylphosphatidylethanolamine (DOPE)/sphingomyelin/cholesterol in a molar ratio of 35:30:15:20 at pH 7.4 versus pH 5. In doing so, we test the hypothesis that fusion of this lipid mixture should be influenced by differences in hydration of DOPE at these two pH values. An examination of the literature reveals that DOPE should be less hydrated at pH 5 (where influenza virus particles fuse with endosome membranes) than at pH 7.4 (where synaptic vesicles or HIV virus particles fuse with plasma membrane). Ensemble kinetic experiments revealed substantial differences in fusion of this plasma membrane mimetic system at these two pH values. The most dramatic difference was the observation of two intermediates at pH 5 but loss of one of these fusion intermediates at pH 7.4. Analysis of data collected at several temperatures also revealed that formation of the initial fusion intermediate (stalk) was favored at pH 7.4 due to increased activation entropy. Our observations support the hypothesis that the different negative intrinsic curvature of DOPE can account for different fusion paths and activation thermodynamics in steps of the fusion process at these two pH values. Finally, the effects of 2 mol % hexadecane on fusion at both pH values seemed to have similar origins for step 1 (promotion of acyl chain or hydrocarbon excursion into interbilayer space) and step 3 (reduction of interstice energy leading to expansion to a critical stalk radius). Different hexadecane effects on activation thermodynamics at these two pH values can also be related to altered DOPE hydration. The results support our kinetic model for fusion and offer insight into the critical role of phosphatidylethanolamine in fusion.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2014 |
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| Erschienen: |
2014 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:107 |
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| Enthalten in: |
Biophysical journal - 107(2014), 6 vom: 16. Sept., Seite 1327-38 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Chakraborty, Hirak [VerfasserIn] |
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| Links: |
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| Anmerkungen: |
Date Completed 20.02.2015 Date Revised 21.10.2021 published: Print Citation Status MEDLINE |
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| doi: |
10.1016/j.bpj.2014.07.048 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| 245 | 1 | 0 | |a pH Alters PEG-mediated fusion of phosphatidylethanolamine-containing vesicles |
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| 520 | |a Copyright © 2014 Biophysical Society. Published by Elsevier Inc. All rights reserved. | ||
| 520 | |a Here, we examine the different mechanisms of poly(ethylene glycol)-mediated fusion of small unilamellar vesicles composed of dioleoylphosphatidylcholine/dioleoylphosphatidylethanolamine (DOPE)/sphingomyelin/cholesterol in a molar ratio of 35:30:15:20 at pH 7.4 versus pH 5. In doing so, we test the hypothesis that fusion of this lipid mixture should be influenced by differences in hydration of DOPE at these two pH values. An examination of the literature reveals that DOPE should be less hydrated at pH 5 (where influenza virus particles fuse with endosome membranes) than at pH 7.4 (where synaptic vesicles or HIV virus particles fuse with plasma membrane). Ensemble kinetic experiments revealed substantial differences in fusion of this plasma membrane mimetic system at these two pH values. The most dramatic difference was the observation of two intermediates at pH 5 but loss of one of these fusion intermediates at pH 7.4. Analysis of data collected at several temperatures also revealed that formation of the initial fusion intermediate (stalk) was favored at pH 7.4 due to increased activation entropy. Our observations support the hypothesis that the different negative intrinsic curvature of DOPE can account for different fusion paths and activation thermodynamics in steps of the fusion process at these two pH values. Finally, the effects of 2 mol % hexadecane on fusion at both pH values seemed to have similar origins for step 1 (promotion of acyl chain or hydrocarbon excursion into interbilayer space) and step 3 (reduction of interstice energy leading to expansion to a critical stalk radius). Different hexadecane effects on activation thermodynamics at these two pH values can also be related to altered DOPE hydration. The results support our kinetic model for fusion and offer insight into the critical role of phosphatidylethanolamine in fusion | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Research Support, N.I.H., Extramural | |
| 650 | 7 | |a Alkanes |2 NLM | |
| 650 | 7 | |a Phosphatidylcholines |2 NLM | |
| 650 | 7 | |a Phosphatidylethanolamines |2 NLM | |
| 650 | 7 | |a Sphingomyelins |2 NLM | |
| 650 | 7 | |a Unilamellar Liposomes |2 NLM | |
| 650 | 7 | |a phosphatidylethanolamine |2 NLM | |
| 650 | 7 | |a 39382-08-6 |2 NLM | |
| 650 | 7 | |a Polyethylene Glycols |2 NLM | |
| 650 | 7 | |a 3WJQ0SDW1A |2 NLM | |
| 650 | 7 | |a Cholesterol |2 NLM | |
| 650 | 7 | |a 97C5T2UQ7J |2 NLM | |
| 650 | 7 | |a 1,2-oleoylphosphatidylcholine |2 NLM | |
| 650 | 7 | |a EDS2L3ODLV |2 NLM | |
| 650 | 7 | |a n-hexadecane |2 NLM | |
| 650 | 7 | |a F8Z00SHP6Q |2 NLM | |
| 700 | 1 | |a Sengupta, Tanusree |e verfasserin |4 aut | |
| 700 | 1 | |a Lentz, Barry R |e verfasserin |4 aut | |
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