Cargando…
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...
Autores principales: | , , , , , , , , , , , |
---|---|
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 |
_version_ | 1785104012333285376 |
---|---|
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. |
format | Online Article Text |
id | pubmed-10491195 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Cold Spring Harbor Laboratory |
record_format | MEDLINE/PubMed |
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 |
work_keys_str_mv | AT kohduksu optopi3kgeneticcodeexpansionandclickchemistryrevealmechanismsunderlyingreciprocalregulationbetweentrpv1andpi3k AT stratiievskaanastasiia optopi3kgeneticcodeexpansionandclickchemistryrevealmechanismsunderlyingreciprocalregulationbetweentrpv1andpi3k AT janasubhashis optopi3kgeneticcodeexpansionandclickchemistryrevealmechanismsunderlyingreciprocalregulationbetweentrpv1andpi3k AT ottoshaunac optopi3kgeneticcodeexpansionandclickchemistryrevealmechanismsunderlyingreciprocalregulationbetweentrpv1andpi3k AT swansonteresam optopi3kgeneticcodeexpansionandclickchemistryrevealmechanismsunderlyingreciprocalregulationbetweentrpv1andpi3k AT nhimanthony optopi3kgeneticcodeexpansionandclickchemistryrevealmechanismsunderlyingreciprocalregulationbetweentrpv1andpi3k AT carlsonsara optopi3kgeneticcodeexpansionandclickchemistryrevealmechanismsunderlyingreciprocalregulationbetweentrpv1andpi3k AT razamarium optopi3kgeneticcodeexpansionandclickchemistryrevealmechanismsunderlyingreciprocalregulationbetweentrpv1andpi3k AT navesligiaaraujo optopi3kgeneticcodeexpansionandclickchemistryrevealmechanismsunderlyingreciprocalregulationbetweentrpv1andpi3k AT senningericn optopi3kgeneticcodeexpansionandclickchemistryrevealmechanismsunderlyingreciprocalregulationbetweentrpv1andpi3k AT mehlryan optopi3kgeneticcodeexpansionandclickchemistryrevealmechanismsunderlyingreciprocalregulationbetweentrpv1andpi3k AT gordonsharonae optopi3kgeneticcodeexpansionandclickchemistryrevealmechanismsunderlyingreciprocalregulationbetweentrpv1andpi3k |