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Coiled-coil formation of the membrane-fusion K/E peptides viewed by electron paramagnetic resonance
The interaction of the complementary K (Ac-(KIAALKE)(3)-GW-NH(2)) and E (Ac-(EIAALEK)(3)-GY-NH(2)) peptides, components of the zipper of an artificial membrane fusion system (Robson Marsden H. et al. Angew Chemie Int Ed. 2009) is investigated by electron paramagnetic resonance (EPR). By frozen solut...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5774749/ https://www.ncbi.nlm.nih.gov/pubmed/29351320 http://dx.doi.org/10.1371/journal.pone.0191197 |
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author | Kumar, Pravin van Son, Martin Zheng, Tingting Valdink, Dayenne Raap, Jan Kros, Alexander Huber, Martina |
author_facet | Kumar, Pravin van Son, Martin Zheng, Tingting Valdink, Dayenne Raap, Jan Kros, Alexander Huber, Martina |
author_sort | Kumar, Pravin |
collection | PubMed |
description | The interaction of the complementary K (Ac-(KIAALKE)(3)-GW-NH(2)) and E (Ac-(EIAALEK)(3)-GY-NH(2)) peptides, components of the zipper of an artificial membrane fusion system (Robson Marsden H. et al. Angew Chemie Int Ed. 2009) is investigated by electron paramagnetic resonance (EPR). By frozen solution continuous-wave EPR and double electron-electron resonance (DEER), the distance between spin labels attached to the K- and to the E-peptide is measured. Three constructs of spin-labelled K- and E-peptides are used in five combinations for low temperature investigations. The K/E heterodimers are found to be parallel, in agreement with previous studies. Also, K homodimers in parallel orientation were observed, a finding that was not reported before. Comparison to room-temperature, solution EPR shows that the latter method is less specific to detect this peptide-peptide interaction. Combining frozen solution cw-EPR for short distances (1.8 nm to 2.0 nm) and DEER for longer distances thus proves versatile to detect the zipper interaction in membrane fusion. As the methodology can be applied to membrane samples, the approach presented suggests itself for in-situ studies of the complete membrane fusion process, opening up new avenues for the study of membrane fusion. |
format | Online Article Text |
id | pubmed-5774749 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-57747492018-02-05 Coiled-coil formation of the membrane-fusion K/E peptides viewed by electron paramagnetic resonance Kumar, Pravin van Son, Martin Zheng, Tingting Valdink, Dayenne Raap, Jan Kros, Alexander Huber, Martina PLoS One Research Article The interaction of the complementary K (Ac-(KIAALKE)(3)-GW-NH(2)) and E (Ac-(EIAALEK)(3)-GY-NH(2)) peptides, components of the zipper of an artificial membrane fusion system (Robson Marsden H. et al. Angew Chemie Int Ed. 2009) is investigated by electron paramagnetic resonance (EPR). By frozen solution continuous-wave EPR and double electron-electron resonance (DEER), the distance between spin labels attached to the K- and to the E-peptide is measured. Three constructs of spin-labelled K- and E-peptides are used in five combinations for low temperature investigations. The K/E heterodimers are found to be parallel, in agreement with previous studies. Also, K homodimers in parallel orientation were observed, a finding that was not reported before. Comparison to room-temperature, solution EPR shows that the latter method is less specific to detect this peptide-peptide interaction. Combining frozen solution cw-EPR for short distances (1.8 nm to 2.0 nm) and DEER for longer distances thus proves versatile to detect the zipper interaction in membrane fusion. As the methodology can be applied to membrane samples, the approach presented suggests itself for in-situ studies of the complete membrane fusion process, opening up new avenues for the study of membrane fusion. Public Library of Science 2018-01-19 /pmc/articles/PMC5774749/ /pubmed/29351320 http://dx.doi.org/10.1371/journal.pone.0191197 Text en © 2018 Kumar et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
spellingShingle | Research Article Kumar, Pravin van Son, Martin Zheng, Tingting Valdink, Dayenne Raap, Jan Kros, Alexander Huber, Martina Coiled-coil formation of the membrane-fusion K/E peptides viewed by electron paramagnetic resonance |
title | Coiled-coil formation of the membrane-fusion K/E peptides viewed by electron paramagnetic resonance |
title_full | Coiled-coil formation of the membrane-fusion K/E peptides viewed by electron paramagnetic resonance |
title_fullStr | Coiled-coil formation of the membrane-fusion K/E peptides viewed by electron paramagnetic resonance |
title_full_unstemmed | Coiled-coil formation of the membrane-fusion K/E peptides viewed by electron paramagnetic resonance |
title_short | Coiled-coil formation of the membrane-fusion K/E peptides viewed by electron paramagnetic resonance |
title_sort | coiled-coil formation of the membrane-fusion k/e peptides viewed by electron paramagnetic resonance |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5774749/ https://www.ncbi.nlm.nih.gov/pubmed/29351320 http://dx.doi.org/10.1371/journal.pone.0191197 |
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