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Real-time observation of functional specialization among phosphorylation sites in CFTR

Phosphoregulation is ubiquitous in biology. Defining the functional roles of individual phosphorylation sites within a multivalent system remains particularly challenging. We have therefore applied a chemical biology approach to light-control the state of single candidate phosphoserines in the canon...

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Autores principales: Infield, Daniel T., Schene, Miranda E., Fazan, Frederico S., Galles, Grace D., Galpin, Jason D., Ahern, Christopher A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Rockefeller University Press 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9930130/
https://www.ncbi.nlm.nih.gov/pubmed/36695813
http://dx.doi.org/10.1085/jgp.202213216
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author Infield, Daniel T.
Schene, Miranda E.
Fazan, Frederico S.
Galles, Grace D.
Galpin, Jason D.
Ahern, Christopher A.
author_facet Infield, Daniel T.
Schene, Miranda E.
Fazan, Frederico S.
Galles, Grace D.
Galpin, Jason D.
Ahern, Christopher A.
author_sort Infield, Daniel T.
collection PubMed
description Phosphoregulation is ubiquitous in biology. Defining the functional roles of individual phosphorylation sites within a multivalent system remains particularly challenging. We have therefore applied a chemical biology approach to light-control the state of single candidate phosphoserines in the canonical anion channel CFTR while simultaneously measuring channel activity. The data show striking non-equivalency among protein kinase A consensus sites, which vary from <10% to >1,000% changes in channel activity upon phosphorylation. Of note, slow phosphorylation of S813 suggests that this site is rate-limiting to the full activation of CFTR. Further, this approach reveals an unexpected coupling between the phosphorylation of S813 and a nearby site, S795. Overall, these data establish an experimental route to understanding roles of specific phosphoserines within complex phosphoregulatory domains. This strategy may be employed in the study of phosphoregulation of other eukaryotic proteins.
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spelling pubmed-99301302023-02-16 Real-time observation of functional specialization among phosphorylation sites in CFTR Infield, Daniel T. Schene, Miranda E. Fazan, Frederico S. Galles, Grace D. Galpin, Jason D. Ahern, Christopher A. J Gen Physiol Article Phosphoregulation is ubiquitous in biology. Defining the functional roles of individual phosphorylation sites within a multivalent system remains particularly challenging. We have therefore applied a chemical biology approach to light-control the state of single candidate phosphoserines in the canonical anion channel CFTR while simultaneously measuring channel activity. The data show striking non-equivalency among protein kinase A consensus sites, which vary from <10% to >1,000% changes in channel activity upon phosphorylation. Of note, slow phosphorylation of S813 suggests that this site is rate-limiting to the full activation of CFTR. Further, this approach reveals an unexpected coupling between the phosphorylation of S813 and a nearby site, S795. Overall, these data establish an experimental route to understanding roles of specific phosphoserines within complex phosphoregulatory domains. This strategy may be employed in the study of phosphoregulation of other eukaryotic proteins. Rockefeller University Press 2023-01-25 /pmc/articles/PMC9930130/ /pubmed/36695813 http://dx.doi.org/10.1085/jgp.202213216 Text en © 2023 Infield et al. https://creativecommons.org/licenses/by/4.0/This article is available under a Creative Commons License (Attribution 4.0 International, as described at https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Infield, Daniel T.
Schene, Miranda E.
Fazan, Frederico S.
Galles, Grace D.
Galpin, Jason D.
Ahern, Christopher A.
Real-time observation of functional specialization among phosphorylation sites in CFTR
title Real-time observation of functional specialization among phosphorylation sites in CFTR
title_full Real-time observation of functional specialization among phosphorylation sites in CFTR
title_fullStr Real-time observation of functional specialization among phosphorylation sites in CFTR
title_full_unstemmed Real-time observation of functional specialization among phosphorylation sites in CFTR
title_short Real-time observation of functional specialization among phosphorylation sites in CFTR
title_sort real-time observation of functional specialization among phosphorylation sites in cftr
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9930130/
https://www.ncbi.nlm.nih.gov/pubmed/36695813
http://dx.doi.org/10.1085/jgp.202213216
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