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

Daniel T Infield; Miranda E Schene; Frederico S Fazan; Grace D Galles; Jason D Galpin; Christopher A Ahern

doi:10.1085/jgp.202213216

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

Journal article

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

Daniel T Infield, Miranda E Schene, Frederico S Fazan, Grace D Galles, Jason D Galpin and Christopher A Ahern

The Journal of general physiology, Vol.155(4), e202213216

04/03/2023

DOI: 10.1085/jgp.202213216

PMCID: PMC9930130

PMID: 36695813

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Abstract

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.

Phosphorylation

Anions - metabolism

Cystic Fibrosis Transmembrane Conductance Regulator - metabolism

Details

Title: Subtitle: Real-time observation of functional specialization among phosphorylation sites in CFTR
Creators: Daniel T Infield - University of Iowa
Miranda E Schene - University of Iowa
Frederico S Fazan - University of Iowa
Grace D Galles - University of Iowa
Jason D Galpin - University of Iowa
Christopher A Ahern - University of Iowa
Resource Type: Journal article
Publication Details: The Journal of general physiology, Vol.155(4), e202213216
DOI: 10.1085/jgp.202213216
PMID: 36695813
PMCID: PMC9930130
NLM abbreviation: J Gen Physiol
ISSN: 0022-1295
eISSN: 1540-7748
Grant note: F32HL149184 / NIH HHS
Language: English
Date published: 04/03/2023
Academic Unit: Molecular Physiology and Biophysics; Iowa Neuroscience Institute
Record Identifier: 9984363504902771

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