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Optical Control of G-Actin with a Photoswitchable Latrunculin

Actin is one of the most abundant proteins in eukaryotic cells and a key component of the cytoskeleton. A range of small molecules have emerged that interfere with actin dynamics by either binding to polymeric F-actin or monomeric G-actin to stabilize or destabilize filaments or prevent their format...

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Autores principales: Vepřek, Nynke A., Cooper, Madeline H., Laprell, Laura, Yang, Emily Jie-Ning, Folkerts, Sander, Bao, Ruiyang, Oertner, Thomas G., Pon, Liza A., Zuchero, J. Bradley, Trauner, Dirk H.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Cold Spring Harbor Laboratory 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10370057/
https://www.ncbi.nlm.nih.gov/pubmed/37502978
http://dx.doi.org/10.1101/2023.07.17.549222
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author Vepřek, Nynke A.
Cooper, Madeline H.
Laprell, Laura
Yang, Emily Jie-Ning
Folkerts, Sander
Bao, Ruiyang
Oertner, Thomas G.
Pon, Liza A.
Zuchero, J. Bradley
Trauner, Dirk H.
author_facet Vepřek, Nynke A.
Cooper, Madeline H.
Laprell, Laura
Yang, Emily Jie-Ning
Folkerts, Sander
Bao, Ruiyang
Oertner, Thomas G.
Pon, Liza A.
Zuchero, J. Bradley
Trauner, Dirk H.
author_sort Vepřek, Nynke A.
collection PubMed
description Actin is one of the most abundant proteins in eukaryotic cells and a key component of the cytoskeleton. A range of small molecules have emerged that interfere with actin dynamics by either binding to polymeric F-actin or monomeric G-actin to stabilize or destabilize filaments or prevent their formation and growth, respectively. Amongst these, the latrunculins, which bind to G-actin and affect polymerization, are widely used as tools to investigate actin-dependent cellular processes. Here, we report a photoswitchable version of latrunculin, termed opto-latrunculin (OptoLat), which binds to G-actin in a light-dependent fashion and affords optical control over actin polymerization. OptoLat can be activated with 390 – 490 nm pulsed light and rapidly relaxes to the inactive form in the dark. Light activated OptoLat induced depolymerization of F-actin networks in oligodendrocytes and budding yeast, as shown by fluorescence microscopy. Subcellular control of actin dynamics in human cancer cell lines was demonstrated by live cell imaging. Light-activated OptoLat also reduced microglia surveillance in organotypic mouse brain slices while ramification was not affected. Incubation in the dark did not alter the structural and functional integrity of microglia. Together, our data demonstrate that OptoLat is a useful tool for the elucidation of G-actin dependent dynamic processes in cells and tissues.
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spelling pubmed-103700572023-07-27 Optical Control of G-Actin with a Photoswitchable Latrunculin Vepřek, Nynke A. Cooper, Madeline H. Laprell, Laura Yang, Emily Jie-Ning Folkerts, Sander Bao, Ruiyang Oertner, Thomas G. Pon, Liza A. Zuchero, J. Bradley Trauner, Dirk H. bioRxiv Article Actin is one of the most abundant proteins in eukaryotic cells and a key component of the cytoskeleton. A range of small molecules have emerged that interfere with actin dynamics by either binding to polymeric F-actin or monomeric G-actin to stabilize or destabilize filaments or prevent their formation and growth, respectively. Amongst these, the latrunculins, which bind to G-actin and affect polymerization, are widely used as tools to investigate actin-dependent cellular processes. Here, we report a photoswitchable version of latrunculin, termed opto-latrunculin (OptoLat), which binds to G-actin in a light-dependent fashion and affords optical control over actin polymerization. OptoLat can be activated with 390 – 490 nm pulsed light and rapidly relaxes to the inactive form in the dark. Light activated OptoLat induced depolymerization of F-actin networks in oligodendrocytes and budding yeast, as shown by fluorescence microscopy. Subcellular control of actin dynamics in human cancer cell lines was demonstrated by live cell imaging. Light-activated OptoLat also reduced microglia surveillance in organotypic mouse brain slices while ramification was not affected. Incubation in the dark did not alter the structural and functional integrity of microglia. Together, our data demonstrate that OptoLat is a useful tool for the elucidation of G-actin dependent dynamic processes in cells and tissues. Cold Spring Harbor Laboratory 2023-07-19 /pmc/articles/PMC10370057/ /pubmed/37502978 http://dx.doi.org/10.1101/2023.07.17.549222 Text en https://creativecommons.org/licenses/by-nc-nd/4.0/This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License (https://creativecommons.org/licenses/by-nc-nd/4.0/) , which allows reusers to copy and distribute the material in any medium or format in unadapted form only, for noncommercial purposes only, and only so long as attribution is given to the creator.
spellingShingle Article
Vepřek, Nynke A.
Cooper, Madeline H.
Laprell, Laura
Yang, Emily Jie-Ning
Folkerts, Sander
Bao, Ruiyang
Oertner, Thomas G.
Pon, Liza A.
Zuchero, J. Bradley
Trauner, Dirk H.
Optical Control of G-Actin with a Photoswitchable Latrunculin
title Optical Control of G-Actin with a Photoswitchable Latrunculin
title_full Optical Control of G-Actin with a Photoswitchable Latrunculin
title_fullStr Optical Control of G-Actin with a Photoswitchable Latrunculin
title_full_unstemmed Optical Control of G-Actin with a Photoswitchable Latrunculin
title_short Optical Control of G-Actin with a Photoswitchable Latrunculin
title_sort optical control of g-actin with a photoswitchable latrunculin
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10370057/
https://www.ncbi.nlm.nih.gov/pubmed/37502978
http://dx.doi.org/10.1101/2023.07.17.549222
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