Modulation of human mitochondrial voltage-dependent anion channel 2 (hVDAC-2) structural stability by cysteine-assisted barrel-lipid interactions
Human mitochondrial voltage-dependent anion channel 2 (hVDAC-2), the most predominant isoform seen in brain mitochondria, is not only crucial for cell survival but is also implicated in Alzheimer disease. The abundance of cysteines in this isoform is particularly fascinating, as hVDAC-1 cysteines have no associated functional role. We report a detailed biophysical examination of a Cys-less mutant of hVDAC-2, and its behavioral comparison with the wild type protein. Our findings suggest that cysteine mutation results in the formation of a better barrel at the expense of weakened protein-lipid interactions. The wild type protein displays stronger lipid association, despite being less structured. A reversal in behavior of both proteins is observed in the case of chemical denaturation, with the Cys-less mutant exhibiting lowered unfolding free energies. In bicellar systems comprising 14-C phosphocholines, we observe that protein-lipid interactions are weakened in both constructs, resulting in barrel structure destabilization. Our biochemical and biophysical studies together reveal key structural roles for the cysteine residues. We find that minor conformational variations in local residues are sufficient to define the membrane protein dynamics in hVDAC-2. Such subtle sequence variations contribute to differential stability of VDACs and may have implications in their in vivo regulation and recycling.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2013 |
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| Erschienen: |
2013 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:288 |
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| Enthalten in: |
The Journal of biological chemistry - 288(2013), 35 vom: 30. Aug., Seite 25584-25592 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Maurya, Svetlana Rajkumar [VerfasserIn] |
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| Links: |
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| Anmerkungen: |
Date Completed 10.12.2013 Date Revised 21.10.2021 published: Print-Electronic Citation Status MEDLINE |
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| doi: |
10.1074/jbc.M113.493692 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| PPN (Katalog-ID): |
NLM229344771 |
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| 100 | 1 | |a Maurya, Svetlana Rajkumar |e verfasserin |4 aut | |
| 245 | 1 | 0 | |a Modulation of human mitochondrial voltage-dependent anion channel 2 (hVDAC-2) structural stability by cysteine-assisted barrel-lipid interactions |
| 264 | 1 | |c 2013 | |
| 336 | |a Text |b txt |2 rdacontent | ||
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| 500 | |a Date Completed 10.12.2013 | ||
| 500 | |a Date Revised 21.10.2021 | ||
| 500 | |a published: Print-Electronic | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a Human mitochondrial voltage-dependent anion channel 2 (hVDAC-2), the most predominant isoform seen in brain mitochondria, is not only crucial for cell survival but is also implicated in Alzheimer disease. The abundance of cysteines in this isoform is particularly fascinating, as hVDAC-1 cysteines have no associated functional role. We report a detailed biophysical examination of a Cys-less mutant of hVDAC-2, and its behavioral comparison with the wild type protein. Our findings suggest that cysteine mutation results in the formation of a better barrel at the expense of weakened protein-lipid interactions. The wild type protein displays stronger lipid association, despite being less structured. A reversal in behavior of both proteins is observed in the case of chemical denaturation, with the Cys-less mutant exhibiting lowered unfolding free energies. In bicellar systems comprising 14-C phosphocholines, we observe that protein-lipid interactions are weakened in both constructs, resulting in barrel structure destabilization. Our biochemical and biophysical studies together reveal key structural roles for the cysteine residues. We find that minor conformational variations in local residues are sufficient to define the membrane protein dynamics in hVDAC-2. Such subtle sequence variations contribute to differential stability of VDACs and may have implications in their in vivo regulation and recycling | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Research Support, Non-U.S. Gov't | |
| 650 | 4 | |a Anion Channel | |
| 650 | 4 | |a Cysteine Mutation | |
| 650 | 4 | |a Kinetics | |
| 650 | 4 | |a Membrane Biophysics | |
| 650 | 4 | |a Membrane Proteins | |
| 650 | 4 | |a Protein Denaturation | |
| 650 | 4 | |a Protein Folding | |
| 650 | 4 | |a Protein-Lipid Interaction | |
| 650 | 4 | |a Spectroscopy | |
| 650 | 4 | |a Thermodynamics | |
| 650 | 7 | |a VDAC2 protein, human |2 NLM | |
| 650 | 7 | |a Voltage-Dependent Anion Channel 2 |2 NLM | |
| 650 | 7 | |a Phosphorylcholine |2 NLM | |
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| 650 | 7 | |a Cysteine |2 NLM | |
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| 700 | 1 | |a Mahalakshmi, Radhakrishnan |e verfasserin |4 aut | |
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| 773 | 1 | 8 | |g volume:288 |g year:2013 |g number:35 |g day:30 |g month:08 |g pages:25584-25592 |
| 856 | 4 | 0 | |u http://dx.doi.org/10.1074/jbc.M113.493692 |3 Volltext |
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