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Spatiotemporally co...
Spatiotemporally controlled generation of NTPs for single-molecule studies
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- Sabantsev, Anton (author)
- Uppsala universitet,Molekylär systembiologi,Science for Life Laboratory, SciLifeLab
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- Mao, Guanzhong, 1985- (author)
- Uppsala universitet,Science for Life Laboratory, SciLifeLab,Molekylär systembiologi
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- Aguirre, Javier Rivera, 1989- (author)
- Uppsala universitet,Molekylär systembiologi,Science for Life Laboratory, SciLifeLab
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- Panfilov, Mikhail (author)
- Peter Great St Petersburg Polytech Univ, St Petersburg, Russia.
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- Arseniev, Anatolii (author)
- Peter Great St Petersburg Polytech Univ, St Petersburg, Russia.;Russian Acad Sci, Ctr Precis Genome Editing & Genet Technol Biomed, Inst Gene Biol, Moscow, Russia.
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- Ho, Oanh (author)
- Uppsala Univ, Dept Cell & Mol Biol, Sci Life Lab, Uppsala, Sweden.
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- Khodorkovskiy, Mikhail (author)
- Peter Great St Petersburg Polytech Univ, St Petersburg, Russia.
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- Deindl, Sebastian (author)
- Uppsala universitet,Molekylär systembiologi,Science for Life Laboratory, SciLifeLab
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(creator_code:org_t)
- 2022-09-21
- 2022
- English.
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In: Nature Chemical Biology. - : Springer Nature. - 1552-4450 .- 1552-4469. ; 18:10, s. 1144-
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Abstract
Subject headings
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- Many essential processes in the cell depend on proteins that use nucleoside triphosphates (NTPs). Methods that directly monitor the often-complex dynamics of these proteins at the single-molecule level have helped to uncover their mechanisms of action. However, the measurement throughput is typically limited for NTP-utilizing reactions, and the quantitative dissection of complex dynamics over multiple sequential turnovers remains challenging. Here we present a method for controlling NTP-driven reactions in single-molecule experiments via the local generation of NTPs (LAGOON) that markedly increases the measurement throughput and enables single-turnover observations. We demonstrate the effectiveness of LAGOON in single-molecule fluorescence and force spectroscopy assays by monitoring DNA unwinding, nucleosome sliding and RNA polymerase elongation. LAGOON can be readily integrated with many single-molecule techniques, and we anticipate that it will facilitate studies of a wide range of crucial NTP-driven processes.
Subject headings
- NATURVETENSKAP -- Biologi -- Biokemi och molekylärbiologi (hsv//swe)
- NATURAL SCIENCES -- Biological Sciences -- Biochemistry and Molecular Biology (hsv//eng)
Publication and Content Type
- ref (subject category)
- art (subject category)
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