Single-molecule thiol-disulfide interchange
<p>Thiol-disulfide interchange occurs widely in cellular processes including redox sensing and homeostasis, protein folding, cell signalling, and the regulation of apoptosis. Nature exploits this chemistry in a highly selective manner, which is challenging to recapitulate in vitro. In this...
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2018
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author | Qing, Y |
author2 | Bayley, J |
author_facet | Bayley, J Qing, Y |
author_sort | Qing, Y |
collection | OXFORD |
description | <p>Thiol-disulfide interchange occurs widely in cellular processes including redox sensing and homeostasis, protein folding, cell signalling, and the regulation of apoptosis. Nature exploits this chemistry in a highly selective manner, which is challenging to recapitulate in vitro. In this thesis, we report site selective and regioselective thioldisulfide interchange on macromolecular disulfide substrates elongated within a protein nanoreactor, where they react with cysteine thiolates presented at different locations along the length of a β strand. Numerous individual reaction events were detected by promoting the substrate turnover. For each substrate, we defined the most reactive cysteines on the β strand and which sulfur atom in the disulfide was attacked and found that the chemistry can be controlled with atomic precision.</p> <p>We further applied our control over the selectivity of thiol-disulfide interchange to the development of a molecular machine. Intrigued by technological potential, scientists have long attempted to control molecular motion. We monitor the individual 0.7-nm steps of a single molecular hopper as it moves in an electric field along a track in a nanopore controlled by a chemical ratchet based on thiol-disulfide interchange. The hopper demonstrates characteristics desired in a moving molecule: defined start- and end-points, processivity, fuel autonomy, directional motion and external control. The hopper is readily functionalized to carry cargos. For example, DNA can be ratcheted along the track in either direction, a prerequisite for nanopore sequencing.</p> |
first_indexed | 2024-03-07T07:05:17Z |
format | Thesis |
id | oxford-uuid:c6ece0e7-4135-4ddc-86cf-7cc9bc0c3b0a |
institution | University of Oxford |
last_indexed | 2024-03-07T07:05:17Z |
publishDate | 2018 |
record_format | dspace |
spelling | oxford-uuid:c6ece0e7-4135-4ddc-86cf-7cc9bc0c3b0a2022-04-25T11:46:01ZSingle-molecule thiol-disulfide interchangeThesishttp://purl.org/coar/resource_type/c_db06uuid:c6ece0e7-4135-4ddc-86cf-7cc9bc0c3b0aORA Deposit2018Qing, YBayley, J<p>Thiol-disulfide interchange occurs widely in cellular processes including redox sensing and homeostasis, protein folding, cell signalling, and the regulation of apoptosis. Nature exploits this chemistry in a highly selective manner, which is challenging to recapitulate in vitro. In this thesis, we report site selective and regioselective thioldisulfide interchange on macromolecular disulfide substrates elongated within a protein nanoreactor, where they react with cysteine thiolates presented at different locations along the length of a β strand. Numerous individual reaction events were detected by promoting the substrate turnover. For each substrate, we defined the most reactive cysteines on the β strand and which sulfur atom in the disulfide was attacked and found that the chemistry can be controlled with atomic precision.</p> <p>We further applied our control over the selectivity of thiol-disulfide interchange to the development of a molecular machine. Intrigued by technological potential, scientists have long attempted to control molecular motion. We monitor the individual 0.7-nm steps of a single molecular hopper as it moves in an electric field along a track in a nanopore controlled by a chemical ratchet based on thiol-disulfide interchange. The hopper demonstrates characteristics desired in a moving molecule: defined start- and end-points, processivity, fuel autonomy, directional motion and external control. The hopper is readily functionalized to carry cargos. For example, DNA can be ratcheted along the track in either direction, a prerequisite for nanopore sequencing.</p> |
spellingShingle | Qing, Y Single-molecule thiol-disulfide interchange |
title | Single-molecule thiol-disulfide interchange |
title_full | Single-molecule thiol-disulfide interchange |
title_fullStr | Single-molecule thiol-disulfide interchange |
title_full_unstemmed | Single-molecule thiol-disulfide interchange |
title_short | Single-molecule thiol-disulfide interchange |
title_sort | single molecule thiol disulfide interchange |
work_keys_str_mv | AT qingy singlemoleculethioldisulfideinterchange |