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OptoPI3K, genetic code expansion, and click chemistry reveal mechanisms underlying reciprocal regulation between TRPV1 and PI3K

Receptor tyrosine kinase signaling is characterized by complex webs of interconnected pathways that regulate diverse cellular functions. The complexity of signaling is a barrier to understanding the pathways that control any particular function. In this work, we use a novel combination of approaches...

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Autores principales: Koh, Duk-Su, Stratiievska, Anastasiia, Jana, Subhashis, Otto, Shauna C., Swanson, Teresa M., Nhim, Anthony, Carlson, Sara, Raza, Marium, Naves, Lígia Araujo, Senning, Eric N., Mehl, Ryan, Gordon, Sharona E.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Cold Spring Harbor Laboratory 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10491195/
https://www.ncbi.nlm.nih.gov/pubmed/37693391
http://dx.doi.org/10.1101/2023.08.29.555449
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author Koh, Duk-Su
Stratiievska, Anastasiia
Jana, Subhashis
Otto, Shauna C.
Swanson, Teresa M.
Nhim, Anthony
Carlson, Sara
Raza, Marium
Naves, Lígia Araujo
Senning, Eric N.
Mehl, Ryan
Gordon, Sharona E.
author_facet Koh, Duk-Su
Stratiievska, Anastasiia
Jana, Subhashis
Otto, Shauna C.
Swanson, Teresa M.
Nhim, Anthony
Carlson, Sara
Raza, Marium
Naves, Lígia Araujo
Senning, Eric N.
Mehl, Ryan
Gordon, Sharona E.
author_sort Koh, Duk-Su
collection PubMed
description Receptor tyrosine kinase signaling is characterized by complex webs of interconnected pathways that regulate diverse cellular functions. The complexity of signaling is a barrier to understanding the pathways that control any particular function. In this work, we use a novel combination of approaches and a new click chemistry probe to determine the role of one pathway in regulating cell surface expression of an ion channel and a receptor tyrosine kinase. We applied an optogenetic approach to uncouple activation of the PI3K pathway from other pathways downstream of RTK activation. In this context, we used genetic code expansion to introduce a click chemistry noncanonical amino acid into the extracellular side of membrane proteins. Applying a cell-impermeant click chemistry fluorophore allowed us to visualize delivery of membrane proteins to the PM in real time. Using these approaches, we demonstrate that activation of PI3K, without activating other pathways downstream of RTK signaling, is sufficient to traffic the TRPV1 ion channels and insulin receptors to the plasma membrane.
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spelling pubmed-104911952023-09-09 OptoPI3K, genetic code expansion, and click chemistry reveal mechanisms underlying reciprocal regulation between TRPV1 and PI3K Koh, Duk-Su Stratiievska, Anastasiia Jana, Subhashis Otto, Shauna C. Swanson, Teresa M. Nhim, Anthony Carlson, Sara Raza, Marium Naves, Lígia Araujo Senning, Eric N. Mehl, Ryan Gordon, Sharona E. bioRxiv Article Receptor tyrosine kinase signaling is characterized by complex webs of interconnected pathways that regulate diverse cellular functions. The complexity of signaling is a barrier to understanding the pathways that control any particular function. In this work, we use a novel combination of approaches and a new click chemistry probe to determine the role of one pathway in regulating cell surface expression of an ion channel and a receptor tyrosine kinase. We applied an optogenetic approach to uncouple activation of the PI3K pathway from other pathways downstream of RTK activation. In this context, we used genetic code expansion to introduce a click chemistry noncanonical amino acid into the extracellular side of membrane proteins. Applying a cell-impermeant click chemistry fluorophore allowed us to visualize delivery of membrane proteins to the PM in real time. Using these approaches, we demonstrate that activation of PI3K, without activating other pathways downstream of RTK signaling, is sufficient to traffic the TRPV1 ion channels and insulin receptors to the plasma membrane. Cold Spring Harbor Laboratory 2023-08-30 /pmc/articles/PMC10491195/ /pubmed/37693391 http://dx.doi.org/10.1101/2023.08.29.555449 Text en https://creativecommons.org/licenses/by/4.0/This work is licensed under a Creative Commons Attribution 4.0 International License (https://creativecommons.org/licenses/by/4.0/) , which allows reusers to distribute, remix, adapt, and build upon the material in any medium or format, so long as attribution is given to the creator. The license allows for commercial use.
spellingShingle Article
Koh, Duk-Su
Stratiievska, Anastasiia
Jana, Subhashis
Otto, Shauna C.
Swanson, Teresa M.
Nhim, Anthony
Carlson, Sara
Raza, Marium
Naves, Lígia Araujo
Senning, Eric N.
Mehl, Ryan
Gordon, Sharona E.
OptoPI3K, genetic code expansion, and click chemistry reveal mechanisms underlying reciprocal regulation between TRPV1 and PI3K
title OptoPI3K, genetic code expansion, and click chemistry reveal mechanisms underlying reciprocal regulation between TRPV1 and PI3K
title_full OptoPI3K, genetic code expansion, and click chemistry reveal mechanisms underlying reciprocal regulation between TRPV1 and PI3K
title_fullStr OptoPI3K, genetic code expansion, and click chemistry reveal mechanisms underlying reciprocal regulation between TRPV1 and PI3K
title_full_unstemmed OptoPI3K, genetic code expansion, and click chemistry reveal mechanisms underlying reciprocal regulation between TRPV1 and PI3K
title_short OptoPI3K, genetic code expansion, and click chemistry reveal mechanisms underlying reciprocal regulation between TRPV1 and PI3K
title_sort optopi3k, genetic code expansion, and click chemistry reveal mechanisms underlying reciprocal regulation between trpv1 and pi3k
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10491195/
https://www.ncbi.nlm.nih.gov/pubmed/37693391
http://dx.doi.org/10.1101/2023.08.29.555449
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