Effect of spin labelling on the aggregation kinetics of yeast prion protein Ure2
Amyloid formation is involved in a wide range of neurodegenerative diseases including Alzheimer's and prion diseases. Structural understanding of the amyloid is critical to delineate the mechanism of aggregation and its pathological spreading. Site-directed spin labelling has emerged as a power...
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| Format: | Article |
| Language: | English |
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The Royal Society
2021-03-01
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| Series: | Royal Society Open Science |
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| Online Access: | https://royalsocietypublishing.org/doi/pdf/10.1098/rsos.201747 |
| _version_ | 1818944698536951808 |
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| author | Emilie N. Liu Giovanna Park Junsuke Nohara Zhefeng Guo |
| author_facet | Emilie N. Liu Giovanna Park Junsuke Nohara Zhefeng Guo |
| author_sort | Emilie N. Liu |
| collection | DOAJ |
| description | Amyloid formation is involved in a wide range of neurodegenerative diseases including Alzheimer's and prion diseases. Structural understanding of the amyloid is critical to delineate the mechanism of aggregation and its pathological spreading. Site-directed spin labelling has emerged as a powerful structural tool in the studies of amyloid structures and provided structural evidence for the parallel in-register β-sheet structure for a wide range of amyloid proteins. It is generally accepted that spin labelling does not disrupt the structure of the amyloid fibrils, the end product of protein aggregation. The effect on the rate of protein aggregation, however, has not been well characterized. Here, we employed a scanning mutagenesis approach to study the effect of spin labelling on the aggregation rate of 79 spin-labelled variants of the Ure2 prion domain. The aggregation of Ure2 protein is the basis of yeast prion [URE3]. We found that all spin-labelled Ure2 mutants aggregated within the experimental timeframe of 15 to 40 h. Among the 79 spin-labelled positions, only five residue sites (N23, N27, S33, I35 and G42) showed a dramatic delay in the aggregation rate as a result of spin labelling. These positions may be important for fibril nucleation, a rate-limiting step in aggregation. Importantly, spin labelling at most of the sites had a muted effect on Ure2 aggregation kinetics, showing a general tolerance of spin labelling in protein aggregation studies. |
| first_indexed | 2024-12-20T07:47:21Z |
| format | Article |
| id | doaj.art-e6862e0feff14956b165eaa12cbe30d3 |
| institution | Directory Open Access Journal |
| issn | 2054-5703 |
| language | English |
| last_indexed | 2024-12-20T07:47:21Z |
| publishDate | 2021-03-01 |
| publisher | The Royal Society |
| record_format | Article |
| series | Royal Society Open Science |
| spelling | doaj.art-e6862e0feff14956b165eaa12cbe30d32022-12-21T19:47:57ZengThe Royal SocietyRoyal Society Open Science2054-57032021-03-018310.1098/rsos.201747201747Effect of spin labelling on the aggregation kinetics of yeast prion protein Ure2Emilie N. LiuGiovanna ParkJunsuke NoharaZhefeng GuoAmyloid formation is involved in a wide range of neurodegenerative diseases including Alzheimer's and prion diseases. Structural understanding of the amyloid is critical to delineate the mechanism of aggregation and its pathological spreading. Site-directed spin labelling has emerged as a powerful structural tool in the studies of amyloid structures and provided structural evidence for the parallel in-register β-sheet structure for a wide range of amyloid proteins. It is generally accepted that spin labelling does not disrupt the structure of the amyloid fibrils, the end product of protein aggregation. The effect on the rate of protein aggregation, however, has not been well characterized. Here, we employed a scanning mutagenesis approach to study the effect of spin labelling on the aggregation rate of 79 spin-labelled variants of the Ure2 prion domain. The aggregation of Ure2 protein is the basis of yeast prion [URE3]. We found that all spin-labelled Ure2 mutants aggregated within the experimental timeframe of 15 to 40 h. Among the 79 spin-labelled positions, only five residue sites (N23, N27, S33, I35 and G42) showed a dramatic delay in the aggregation rate as a result of spin labelling. These positions may be important for fibril nucleation, a rate-limiting step in aggregation. Importantly, spin labelling at most of the sites had a muted effect on Ure2 aggregation kinetics, showing a general tolerance of spin labelling in protein aggregation studies.https://royalsocietypublishing.org/doi/pdf/10.1098/rsos.201747spin labellingamyloidprotein aggregationyeast prionelectron paramagnetic resonance |
| spellingShingle | Emilie N. Liu Giovanna Park Junsuke Nohara Zhefeng Guo Effect of spin labelling on the aggregation kinetics of yeast prion protein Ure2 Royal Society Open Science spin labelling amyloid protein aggregation yeast prion electron paramagnetic resonance |
| title | Effect of spin labelling on the aggregation kinetics of yeast prion protein Ure2 |
| title_full | Effect of spin labelling on the aggregation kinetics of yeast prion protein Ure2 |
| title_fullStr | Effect of spin labelling on the aggregation kinetics of yeast prion protein Ure2 |
| title_full_unstemmed | Effect of spin labelling on the aggregation kinetics of yeast prion protein Ure2 |
| title_short | Effect of spin labelling on the aggregation kinetics of yeast prion protein Ure2 |
| title_sort | effect of spin labelling on the aggregation kinetics of yeast prion protein ure2 |
| topic | spin labelling amyloid protein aggregation yeast prion electron paramagnetic resonance |
| url | https://royalsocietypublishing.org/doi/pdf/10.1098/rsos.201747 |
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