Monitoring site-specific conformational changes in real-time reveals a misfolding mechanism of the prion protein
During pathological aggregation, proteins undergo remarkable conformational re-arrangements to anomalously assemble into a heterogeneous collection of misfolded multimers, ranging from soluble oligomers to insoluble amyloid fibrils. Inspired by fluorescence resonance energy transfer (FRET) measureme...
| Main Authors: | , |
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| Format: | Article |
| Language: | English |
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eLife Sciences Publications Ltd
2019-06-01
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| Series: | eLife |
| Subjects: | |
| Online Access: | https://elifesciences.org/articles/44698 |
| _version_ | 1811253015786553344 |
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| author | Ishita Sengupta Jayant Udgaonkar |
| author_facet | Ishita Sengupta Jayant Udgaonkar |
| author_sort | Ishita Sengupta |
| collection | DOAJ |
| description | During pathological aggregation, proteins undergo remarkable conformational re-arrangements to anomalously assemble into a heterogeneous collection of misfolded multimers, ranging from soluble oligomers to insoluble amyloid fibrils. Inspired by fluorescence resonance energy transfer (FRET) measurements of protein folding, an experimental strategy to study site-specific misfolding kinetics during aggregation, by effectively suppressing contributions from inter-molecular FRET, is described. Specifically, the kinetics of conformational changes across different secondary and tertiary structural segments of the mouse prion protein (moPrP) were monitored independently, after the monomeric units transformed into large oligomers OL, which subsequently disaggregated reversibly into small oligomers OS at pH 4. The sequence segments spanning helices α2 and α3 underwent a compaction during the formation of OL and elongation into β-sheets during the formation of OS. The β1-α1-β2 and α2-α3 subdomains were separated, and the helix α1 was unfolded to varying extents in both OL and OS. |
| first_indexed | 2024-04-12T16:43:22Z |
| format | Article |
| id | doaj.art-71b9195744174abf92fe2944c312c26b |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-04-12T16:43:22Z |
| publishDate | 2019-06-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-71b9195744174abf92fe2944c312c26b2022-12-22T03:24:40ZengeLife Sciences Publications LtdeLife2050-084X2019-06-01810.7554/eLife.44698Monitoring site-specific conformational changes in real-time reveals a misfolding mechanism of the prion proteinIshita Sengupta0https://orcid.org/0000-0002-2679-6954Jayant Udgaonkar1https://orcid.org/0000-0002-7005-224XNational Centre for Biological Sciences, Tata Institute of Fundamental Research, Bengaluru, IndiaNational Centre for Biological Sciences, Tata Institute of Fundamental Research, Bengaluru, IndiaDuring pathological aggregation, proteins undergo remarkable conformational re-arrangements to anomalously assemble into a heterogeneous collection of misfolded multimers, ranging from soluble oligomers to insoluble amyloid fibrils. Inspired by fluorescence resonance energy transfer (FRET) measurements of protein folding, an experimental strategy to study site-specific misfolding kinetics during aggregation, by effectively suppressing contributions from inter-molecular FRET, is described. Specifically, the kinetics of conformational changes across different secondary and tertiary structural segments of the mouse prion protein (moPrP) were monitored independently, after the monomeric units transformed into large oligomers OL, which subsequently disaggregated reversibly into small oligomers OS at pH 4. The sequence segments spanning helices α2 and α3 underwent a compaction during the formation of OL and elongation into β-sheets during the formation of OS. The β1-α1-β2 and α2-α3 subdomains were separated, and the helix α1 was unfolded to varying extents in both OL and OS.https://elifesciences.org/articles/44698prionα to β switchintra-molecular FRETsmall and large oligomers |
| spellingShingle | Ishita Sengupta Jayant Udgaonkar Monitoring site-specific conformational changes in real-time reveals a misfolding mechanism of the prion protein eLife prion α to β switch intra-molecular FRET small and large oligomers |
| title | Monitoring site-specific conformational changes in real-time reveals a misfolding mechanism of the prion protein |
| title_full | Monitoring site-specific conformational changes in real-time reveals a misfolding mechanism of the prion protein |
| title_fullStr | Monitoring site-specific conformational changes in real-time reveals a misfolding mechanism of the prion protein |
| title_full_unstemmed | Monitoring site-specific conformational changes in real-time reveals a misfolding mechanism of the prion protein |
| title_short | Monitoring site-specific conformational changes in real-time reveals a misfolding mechanism of the prion protein |
| title_sort | monitoring site specific conformational changes in real time reveals a misfolding mechanism of the prion protein |
| topic | prion α to β switch intra-molecular FRET small and large oligomers |
| url | https://elifesciences.org/articles/44698 |
| work_keys_str_mv | AT ishitasengupta monitoringsitespecificconformationalchangesinrealtimerevealsamisfoldingmechanismoftheprionprotein AT jayantudgaonkar monitoringsitespecificconformationalchangesinrealtimerevealsamisfoldingmechanismoftheprionprotein |