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Single-molecule nanopore sensing of actin dynamics and drug binding

Actin is a key protein in the dynamic processes within the eukaryotic cell. To date, methods exploring the molecular state of actin are limited to insights gained from structural approaches, providing a snapshot of protein folding, or methods that require chemical modifications compromising actin mo...

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Autores principales: Wang, Xiaoyi, Wilkinson, Mark D., Lin, Xiaoyan, Ren, Ren, Willison, Keith R., Ivanov, Aleksandar P., Baum, Jake, Edel, Joshua B.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The Royal Society of Chemistry 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8146688/
https://www.ncbi.nlm.nih.gov/pubmed/34084351
http://dx.doi.org/10.1039/c9sc05710b
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author Wang, Xiaoyi
Wilkinson, Mark D.
Lin, Xiaoyan
Ren, Ren
Willison, Keith R.
Ivanov, Aleksandar P.
Baum, Jake
Edel, Joshua B.
author_facet Wang, Xiaoyi
Wilkinson, Mark D.
Lin, Xiaoyan
Ren, Ren
Willison, Keith R.
Ivanov, Aleksandar P.
Baum, Jake
Edel, Joshua B.
author_sort Wang, Xiaoyi
collection PubMed
description Actin is a key protein in the dynamic processes within the eukaryotic cell. To date, methods exploring the molecular state of actin are limited to insights gained from structural approaches, providing a snapshot of protein folding, or methods that require chemical modifications compromising actin monomer thermostability. Nanopore sensing permits label-free investigation of native proteins and is ideally suited to study proteins such as actin that require specialised buffers and cofactors. Using nanopores, we determined the state of actin at the macromolecular level (filamentous or globular) and in its monomeric form bound to inhibitors. We revealed urea-dependent and voltage-dependent transitional states and observed the unfolding process within which sub-populations of transient actin oligomers are visible. We detected, in real-time, filament-growth, and drug-binding at the single-molecule level demonstrating the promise of nanopore sensing for in-depth understanding of protein folding landscapes and for drug discovery.
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spelling pubmed-81466882021-06-02 Single-molecule nanopore sensing of actin dynamics and drug binding Wang, Xiaoyi Wilkinson, Mark D. Lin, Xiaoyan Ren, Ren Willison, Keith R. Ivanov, Aleksandar P. Baum, Jake Edel, Joshua B. Chem Sci Chemistry Actin is a key protein in the dynamic processes within the eukaryotic cell. To date, methods exploring the molecular state of actin are limited to insights gained from structural approaches, providing a snapshot of protein folding, or methods that require chemical modifications compromising actin monomer thermostability. Nanopore sensing permits label-free investigation of native proteins and is ideally suited to study proteins such as actin that require specialised buffers and cofactors. Using nanopores, we determined the state of actin at the macromolecular level (filamentous or globular) and in its monomeric form bound to inhibitors. We revealed urea-dependent and voltage-dependent transitional states and observed the unfolding process within which sub-populations of transient actin oligomers are visible. We detected, in real-time, filament-growth, and drug-binding at the single-molecule level demonstrating the promise of nanopore sensing for in-depth understanding of protein folding landscapes and for drug discovery. The Royal Society of Chemistry 2019-12-03 /pmc/articles/PMC8146688/ /pubmed/34084351 http://dx.doi.org/10.1039/c9sc05710b Text en This journal is © The Royal Society of Chemistry https://creativecommons.org/licenses/by/3.0/
spellingShingle Chemistry
Wang, Xiaoyi
Wilkinson, Mark D.
Lin, Xiaoyan
Ren, Ren
Willison, Keith R.
Ivanov, Aleksandar P.
Baum, Jake
Edel, Joshua B.
Single-molecule nanopore sensing of actin dynamics and drug binding
title Single-molecule nanopore sensing of actin dynamics and drug binding
title_full Single-molecule nanopore sensing of actin dynamics and drug binding
title_fullStr Single-molecule nanopore sensing of actin dynamics and drug binding
title_full_unstemmed Single-molecule nanopore sensing of actin dynamics and drug binding
title_short Single-molecule nanopore sensing of actin dynamics and drug binding
title_sort single-molecule nanopore sensing of actin dynamics and drug binding
topic Chemistry
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8146688/
https://www.ncbi.nlm.nih.gov/pubmed/34084351
http://dx.doi.org/10.1039/c9sc05710b
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