Coiled-coil heterodimers with increased stability for cellular regulation and sensing SARS-CoV-2 spike protein-mediated cell fusion
Abstract Coiled-coil (CC) dimer-forming peptides are attractive designable modules for mediating protein association. Highly stable CCs are desired for biological activity regulation and assay. Here, we report the design and versatile applications of orthogonal CC dimer-forming peptides with a disso...
Guardado en:
| Autores principales: | , , , , , , |
|---|---|
| Formato: | article |
| Lenguaje: | EN |
| Publicado: |
Nature Portfolio
2021
|
| Materias: | |
| Acceso en línea: | https://doaj.org/article/24366d44f44440bd9d30fddabfebcaae |
| Etiquetas: |
Agregar Etiqueta
Sin Etiquetas, Sea el primero en etiquetar este registro!
|
| id |
oai:doaj.org-article:24366d44f44440bd9d30fddabfebcaae |
|---|---|
| record_format |
dspace |
| spelling |
oai:doaj.org-article:24366d44f44440bd9d30fddabfebcaae2021-12-02T17:39:31ZCoiled-coil heterodimers with increased stability for cellular regulation and sensing SARS-CoV-2 spike protein-mediated cell fusion10.1038/s41598-021-88315-32045-2322https://doaj.org/article/24366d44f44440bd9d30fddabfebcaae2021-04-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-88315-3https://doaj.org/toc/2045-2322Abstract Coiled-coil (CC) dimer-forming peptides are attractive designable modules for mediating protein association. Highly stable CCs are desired for biological activity regulation and assay. Here, we report the design and versatile applications of orthogonal CC dimer-forming peptides with a dissociation constant in the low nanomolar range. In vitro stability and specificity was confirmed in mammalian cells by enzyme reconstitution, transcriptional activation using a combination of DNA-binding and a transcriptional activation domain, and cellular-enzyme-activity regulation based on externally-added peptides. In addition to cellular regulation, coiled-coil-mediated reporter reconstitution was used for the detection of cell fusion mediated by the interaction between the spike protein of pandemic SARS-CoV2 and the ACE2 receptor. This assay can be used to investigate the mechanism of viral spike protein-mediated fusion or screening for viral inhibitors under biosafety level 1 conditions.Tjaša PlaperJana AupičPetra DeklevaFabio LapentaMateja Manček KeberRoman JeralaMojca BenčinaNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-16 (2021) |
| institution |
DOAJ |
| collection |
DOAJ |
| language |
EN |
| topic |
Medicine R Science Q |
| spellingShingle |
Medicine R Science Q Tjaša Plaper Jana Aupič Petra Dekleva Fabio Lapenta Mateja Manček Keber Roman Jerala Mojca Benčina Coiled-coil heterodimers with increased stability for cellular regulation and sensing SARS-CoV-2 spike protein-mediated cell fusion |
| description |
Abstract Coiled-coil (CC) dimer-forming peptides are attractive designable modules for mediating protein association. Highly stable CCs are desired for biological activity regulation and assay. Here, we report the design and versatile applications of orthogonal CC dimer-forming peptides with a dissociation constant in the low nanomolar range. In vitro stability and specificity was confirmed in mammalian cells by enzyme reconstitution, transcriptional activation using a combination of DNA-binding and a transcriptional activation domain, and cellular-enzyme-activity regulation based on externally-added peptides. In addition to cellular regulation, coiled-coil-mediated reporter reconstitution was used for the detection of cell fusion mediated by the interaction between the spike protein of pandemic SARS-CoV2 and the ACE2 receptor. This assay can be used to investigate the mechanism of viral spike protein-mediated fusion or screening for viral inhibitors under biosafety level 1 conditions. |
| format |
article |
| author |
Tjaša Plaper Jana Aupič Petra Dekleva Fabio Lapenta Mateja Manček Keber Roman Jerala Mojca Benčina |
| author_facet |
Tjaša Plaper Jana Aupič Petra Dekleva Fabio Lapenta Mateja Manček Keber Roman Jerala Mojca Benčina |
| author_sort |
Tjaša Plaper |
| title |
Coiled-coil heterodimers with increased stability for cellular regulation and sensing SARS-CoV-2 spike protein-mediated cell fusion |
| title_short |
Coiled-coil heterodimers with increased stability for cellular regulation and sensing SARS-CoV-2 spike protein-mediated cell fusion |
| title_full |
Coiled-coil heterodimers with increased stability for cellular regulation and sensing SARS-CoV-2 spike protein-mediated cell fusion |
| title_fullStr |
Coiled-coil heterodimers with increased stability for cellular regulation and sensing SARS-CoV-2 spike protein-mediated cell fusion |
| title_full_unstemmed |
Coiled-coil heterodimers with increased stability for cellular regulation and sensing SARS-CoV-2 spike protein-mediated cell fusion |
| title_sort |
coiled-coil heterodimers with increased stability for cellular regulation and sensing sars-cov-2 spike protein-mediated cell fusion |
| publisher |
Nature Portfolio |
| publishDate |
2021 |
| url |
https://doaj.org/article/24366d44f44440bd9d30fddabfebcaae |
| work_keys_str_mv |
AT tjasaplaper coiledcoilheterodimerswithincreasedstabilityforcellularregulationandsensingsarscov2spikeproteinmediatedcellfusion AT janaaupic coiledcoilheterodimerswithincreasedstabilityforcellularregulationandsensingsarscov2spikeproteinmediatedcellfusion AT petradekleva coiledcoilheterodimerswithincreasedstabilityforcellularregulationandsensingsarscov2spikeproteinmediatedcellfusion AT fabiolapenta coiledcoilheterodimerswithincreasedstabilityforcellularregulationandsensingsarscov2spikeproteinmediatedcellfusion AT matejamancekkeber coiledcoilheterodimerswithincreasedstabilityforcellularregulationandsensingsarscov2spikeproteinmediatedcellfusion AT romanjerala coiledcoilheterodimerswithincreasedstabilityforcellularregulationandsensingsarscov2spikeproteinmediatedcellfusion AT mojcabencina coiledcoilheterodimerswithincreasedstabilityforcellularregulationandsensingsarscov2spikeproteinmediatedcellfusion |
| _version_ |
1718379812570005504 |