Monitoring multiple distances within a single molecule using switchable FRET.
The analysis of structure and dynamics of biomolecules is important for understanding their function. Toward this aim, we introduce a method called 'switchable FRET', which combines single-molecule fluorescence resonance energy transfer (FRET) with reversible photoswitching of fluorophores...
| Main Authors: | , , , , , , |
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| Format: | Journal article |
| Language: | English |
| Published: |
2010
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| _version_ | 1797071014749798400 |
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| author | Uphoff, S Holden, S Le Reste, L Periz, J van de Linde, S Heilemann, M Kapanidis, A |
| author_facet | Uphoff, S Holden, S Le Reste, L Periz, J van de Linde, S Heilemann, M Kapanidis, A |
| author_sort | Uphoff, S |
| collection | OXFORD |
| description | The analysis of structure and dynamics of biomolecules is important for understanding their function. Toward this aim, we introduce a method called 'switchable FRET', which combines single-molecule fluorescence resonance energy transfer (FRET) with reversible photoswitching of fluorophores. Typically, single-molecule FRET is measured within a single donor-acceptor pair and reports on only one distance. Although multipair FRET approaches that monitor multiple distances have been developed, they are technically challenging and difficult to extend, mainly because of their reliance on spectrally distinct acceptors. In contrast, switchable FRET sequentially probes FRET between a single donor and spectrally identical photoswitchable acceptors, dramatically reducing the experimental and analytical complexity and enabling direct monitoring of multiple distances. Our experiments on DNA molecules, a protein-DNA complex and dynamic Holliday junctions demonstrate the potential of switchable FRET for studying dynamic, multicomponent biomolecules. |
| first_indexed | 2024-03-06T22:47:10Z |
| format | Journal article |
| id | oxford-uuid:5d8f6cbb-1283-4957-8c55-48a4024bed76 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-06T22:47:10Z |
| publishDate | 2010 |
| record_format | dspace |
| spelling | oxford-uuid:5d8f6cbb-1283-4957-8c55-48a4024bed762022-03-26T17:35:15ZMonitoring multiple distances within a single molecule using switchable FRET.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:5d8f6cbb-1283-4957-8c55-48a4024bed76EnglishSymplectic Elements at Oxford2010Uphoff, SHolden, SLe Reste, LPeriz, Jvan de Linde, SHeilemann, MKapanidis, AThe analysis of structure and dynamics of biomolecules is important for understanding their function. Toward this aim, we introduce a method called 'switchable FRET', which combines single-molecule fluorescence resonance energy transfer (FRET) with reversible photoswitching of fluorophores. Typically, single-molecule FRET is measured within a single donor-acceptor pair and reports on only one distance. Although multipair FRET approaches that monitor multiple distances have been developed, they are technically challenging and difficult to extend, mainly because of their reliance on spectrally distinct acceptors. In contrast, switchable FRET sequentially probes FRET between a single donor and spectrally identical photoswitchable acceptors, dramatically reducing the experimental and analytical complexity and enabling direct monitoring of multiple distances. Our experiments on DNA molecules, a protein-DNA complex and dynamic Holliday junctions demonstrate the potential of switchable FRET for studying dynamic, multicomponent biomolecules. |
| spellingShingle | Uphoff, S Holden, S Le Reste, L Periz, J van de Linde, S Heilemann, M Kapanidis, A Monitoring multiple distances within a single molecule using switchable FRET. |
| title | Monitoring multiple distances within a single molecule using switchable FRET. |
| title_full | Monitoring multiple distances within a single molecule using switchable FRET. |
| title_fullStr | Monitoring multiple distances within a single molecule using switchable FRET. |
| title_full_unstemmed | Monitoring multiple distances within a single molecule using switchable FRET. |
| title_short | Monitoring multiple distances within a single molecule using switchable FRET. |
| title_sort | monitoring multiple distances within a single molecule using switchable fret |
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