Reduced and mutant lysozyme refolding with lipid vesicles. Model study of disulfide impact on equilibria and dynamics
Copyright © 2016 Elsevier B.V. All rights reserved..
The recovery of secondary structure in disordered, disulfide-reduced hen egg white lysozyme (HEWL) upon interaction with lipid vesicles was studied using circular dichroism (CD), fluorescence and infrared (IR) spectroscopic techniques. Lipid vesicles having negative head groups, such as DMPG, interact with reduced HEWL to induce formation of more helical structure than in native HEWL, but no stable tertiary structure was evident. Changes in tertiary structure, as evidenced by local environment of the tryptophan residues, were monitored by fluorescence. Spectra for oxidized HEWL, reduced HEWL and mutants with no or just one disulfide bond developed variable degrees of increased helicity when added to negatively charged lipid vesicles, mostly depending on packing of tails. When mixed with zwitterionic lipid vesicles, reduced HEWL developed β-sheet structure with no change in helicity, indicating an altered interaction mechanism. Stopped flow CD and fluorescence dynamics, were fit to multi-exponential forms, consistent with refolding to metastable intermediates of increasing helicity for HEWL interacting with lipid vesicles. Formation of an intermediate after rapid interaction of the lipid vesicles and the protein is supported by the correlation of faster steps in CD and fluorescence kinetics, and largely appears driven by electrostatic interaction. In subsequent slower steps, the partially refolded intermediate further alters structure, gaining helicity and modifying tryptophan packing, as driven by hydrophobic interactions.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2016 |
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| Erschienen: |
2016 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:1864 |
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| Enthalten in: |
Biochimica et biophysica acta - 1864(2016), 9 vom: 08. Sept., Seite 1083-1092 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Zhu, Weiying [VerfasserIn] |
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| Links: |
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| Anmerkungen: |
Date Completed 04.10.2017 Date Revised 04.10.2017 published: Print-Electronic Citation Status MEDLINE |
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| doi: |
10.1016/j.bbapap.2016.05.010 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| PPN (Katalog-ID): |
NLM260875406 |
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| 100 | 1 | |a Zhu, Weiying |e verfasserin |4 aut | |
| 245 | 1 | 0 | |a Reduced and mutant lysozyme refolding with lipid vesicles. Model study of disulfide impact on equilibria and dynamics |
| 264 | 1 | |c 2016 | |
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| 500 | |a Date Revised 04.10.2017 | ||
| 500 | |a published: Print-Electronic | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a Copyright © 2016 Elsevier B.V. All rights reserved. | ||
| 520 | |a The recovery of secondary structure in disordered, disulfide-reduced hen egg white lysozyme (HEWL) upon interaction with lipid vesicles was studied using circular dichroism (CD), fluorescence and infrared (IR) spectroscopic techniques. Lipid vesicles having negative head groups, such as DMPG, interact with reduced HEWL to induce formation of more helical structure than in native HEWL, but no stable tertiary structure was evident. Changes in tertiary structure, as evidenced by local environment of the tryptophan residues, were monitored by fluorescence. Spectra for oxidized HEWL, reduced HEWL and mutants with no or just one disulfide bond developed variable degrees of increased helicity when added to negatively charged lipid vesicles, mostly depending on packing of tails. When mixed with zwitterionic lipid vesicles, reduced HEWL developed β-sheet structure with no change in helicity, indicating an altered interaction mechanism. Stopped flow CD and fluorescence dynamics, were fit to multi-exponential forms, consistent with refolding to metastable intermediates of increasing helicity for HEWL interacting with lipid vesicles. Formation of an intermediate after rapid interaction of the lipid vesicles and the protein is supported by the correlation of faster steps in CD and fluorescence kinetics, and largely appears driven by electrostatic interaction. In subsequent slower steps, the partially refolded intermediate further alters structure, gaining helicity and modifying tryptophan packing, as driven by hydrophobic interactions | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Research Support, Non-U.S. Gov't | |
| 650 | 4 | |a CD | |
| 650 | 4 | |a Disulfide | |
| 650 | 4 | |a Fluorescence | |
| 650 | 4 | |a IR | |
| 650 | 4 | |a Kinetics | |
| 650 | 4 | |a Lipid vesicles | |
| 650 | 4 | |a Protein folding | |
| 650 | 4 | |a Reduction | |
| 650 | 7 | |a Liposomes |2 NLM | |
| 650 | 7 | |a Phosphatidylglycerols |2 NLM | |
| 650 | 7 | |a 1,2-dioleoyl-sn-glycero-3-phosphoglycerol |2 NLM | |
| 650 | 7 | |a 66322-31-4 |2 NLM | |
| 650 | 7 | |a 1-palmitoyl-2-oleoylglycero-3-phosphoglycerol |2 NLM | |
| 650 | 7 | |a 81490-05-3 |2 NLM | |
| 650 | 7 | |a dimyristoylphosphatidylglycerol |2 NLM | |
| 650 | 7 | |a BI71WT9P3R |2 NLM | |
| 650 | 7 | |a hen egg lysozyme |2 NLM | |
| 650 | 7 | |a EC 3.2.1.- |2 NLM | |
| 650 | 7 | |a Muramidase |2 NLM | |
| 650 | 7 | |a EC 3.2.1.17 |2 NLM | |
| 700 | 1 | |a Silvers, Robert |e verfasserin |4 aut | |
| 700 | 1 | |a Schwalbe, Harald |e verfasserin |4 aut | |
| 700 | 1 | |a Keiderling, Timothy A |e verfasserin |4 aut | |
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