Journal article
Mobile molecules: reactivity profiling guides faster movement on a cysteine track
- Abstract:
- We previously reported a molecular hopper, which makes sub-nanometer steps by thiol-disulfide interchange along a track with cysteine footholds within a protein nanopore. Here we optimize the hopping rate (ca. 0.1 s−1 in the previous work) with a view towards rapid enzymeless biopolymer characterization during translocation within nanopores. We first took a single-molecule approach to obtain the reactivity profiles of individual footholds. The pKa values of cysteine thiols within a pore ranged from 9.17 to 9.85, and the pH-independent rate constants of the thiolates with a small-molecule disulfide varied by up to 20-fold. Through site-specific mutagenesis and a pH increase from 8.5 to 9.5, the overall hopping rate of a DNA cargo along a five-cysteine track was accelerated 4-fold, and the rate-limiting step 21-fold.
- Publication status:
- Published
- Peer review status:
- Peer reviewed
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(Preview, Version of record, pdf, 4.3MB, Terms of use)
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- Publisher copy:
- 10.1002/anie.202300890
Authors
+ European Research Council
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- Funder identifier:
- https://ror.org/0472cxd90
- Grant:
- 833792
- Programme:
- H2020
+ Wellcome Trust
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- Funder identifier:
- https://ror.org/029chgv08
- Programme:
- Wellcome Leap Delta Tissue Program
+ Engineering and Physical Sciences Research Council
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- Funder identifier:
- https://ror.org/0439y7842
- Grant:
- EP/T022205/1
- EP/L015838/1
- Publisher:
- Wiley
- Journal:
- Angewandte Chemie International Edition More from this journal
- Volume:
- 62
- Issue:
- 21
- Article number:
- e202300890
- Place of publication:
- Germany
- Publication date:
- 2023-04-13
- DOI:
- EISSN:
-
1521-3773
- ISSN:
-
1433-7851
- Pmid:
-
36930533
- Language:
-
English
- Keywords:
- Pubs id:
-
1334088
- Local pid:
-
pubs:1334088
- Deposit date:
-
2025-02-28
- ARK identifier:
Terms of use
- Copyright holder:
- Bo et al
- Copyright date:
- 2023
- Rights statement:
- © 2023 The Authors. Angewandte Chemie International Edition published by Wiley-VCH GmbH. This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
- Licence:
- CC Attribution (CC BY)
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