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Chemical genetic identification of GAK substrates reveals its role in regulating Na(+)/K(+)-ATPase

Cyclin G–associated kinase (GAK) is a ubiquitous serine/threonine kinase that facilitates clathrin uncoating during vesicle trafficking. GAK phosphorylates a coat adaptor component, AP2M1, to help achieve this function. GAK is also implicated in Parkinson's disease through genome-wide associati...

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Autores principales: Lin, Amy W, Gill, Kalbinder K, Castañeda, Marisol Sampedro, Matucci, Irene, Eder, Noreen, Claxton, Suzanne, Flynn, Helen, Snijders, Ambrosius P, George, Roger, Ultanir, Sila K
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Life Science Alliance LLC 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6312924/
https://www.ncbi.nlm.nih.gov/pubmed/30623173
http://dx.doi.org/10.26508/lsa.201800118
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author Lin, Amy W
Gill, Kalbinder K
Castañeda, Marisol Sampedro
Matucci, Irene
Eder, Noreen
Claxton, Suzanne
Flynn, Helen
Snijders, Ambrosius P
George, Roger
Ultanir, Sila K
author_facet Lin, Amy W
Gill, Kalbinder K
Castañeda, Marisol Sampedro
Matucci, Irene
Eder, Noreen
Claxton, Suzanne
Flynn, Helen
Snijders, Ambrosius P
George, Roger
Ultanir, Sila K
author_sort Lin, Amy W
collection PubMed
description Cyclin G–associated kinase (GAK) is a ubiquitous serine/threonine kinase that facilitates clathrin uncoating during vesicle trafficking. GAK phosphorylates a coat adaptor component, AP2M1, to help achieve this function. GAK is also implicated in Parkinson's disease through genome-wide association studies. However, GAK's role in mammalian neurons remains unclear, and insight may come from identification of further substrates. Employing a chemical genetics method, we show here that the sodium potassium pump (Na(+)/K(+)-ATPase) α-subunit Atp1a3 is a GAK target and that GAK regulates Na(+)/K(+)-ATPase trafficking to the plasma membrane. Whole-cell patch clamp recordings from CA1 pyramidal neurons in GAK conditional knockout mice show a larger change in resting membrane potential when exposed to the Na(+)/K(+)-ATPase blocker ouabain, indicating compromised Na(+)/K(+)-ATPase function in GAK knockouts. Our results suggest a modulatory role for GAK via phosphoregulation of substrates such as Atp1a3 during cargo trafficking.
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spelling pubmed-63129242019-01-08 Chemical genetic identification of GAK substrates reveals its role in regulating Na(+)/K(+)-ATPase Lin, Amy W Gill, Kalbinder K Castañeda, Marisol Sampedro Matucci, Irene Eder, Noreen Claxton, Suzanne Flynn, Helen Snijders, Ambrosius P George, Roger Ultanir, Sila K Life Sci Alliance Research Articles Cyclin G–associated kinase (GAK) is a ubiquitous serine/threonine kinase that facilitates clathrin uncoating during vesicle trafficking. GAK phosphorylates a coat adaptor component, AP2M1, to help achieve this function. GAK is also implicated in Parkinson's disease through genome-wide association studies. However, GAK's role in mammalian neurons remains unclear, and insight may come from identification of further substrates. Employing a chemical genetics method, we show here that the sodium potassium pump (Na(+)/K(+)-ATPase) α-subunit Atp1a3 is a GAK target and that GAK regulates Na(+)/K(+)-ATPase trafficking to the plasma membrane. Whole-cell patch clamp recordings from CA1 pyramidal neurons in GAK conditional knockout mice show a larger change in resting membrane potential when exposed to the Na(+)/K(+)-ATPase blocker ouabain, indicating compromised Na(+)/K(+)-ATPase function in GAK knockouts. Our results suggest a modulatory role for GAK via phosphoregulation of substrates such as Atp1a3 during cargo trafficking. Life Science Alliance LLC 2018-12-31 /pmc/articles/PMC6312924/ /pubmed/30623173 http://dx.doi.org/10.26508/lsa.201800118 Text en © 2018 Lin 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 Research Articles
Lin, Amy W
Gill, Kalbinder K
Castañeda, Marisol Sampedro
Matucci, Irene
Eder, Noreen
Claxton, Suzanne
Flynn, Helen
Snijders, Ambrosius P
George, Roger
Ultanir, Sila K
Chemical genetic identification of GAK substrates reveals its role in regulating Na(+)/K(+)-ATPase
title Chemical genetic identification of GAK substrates reveals its role in regulating Na(+)/K(+)-ATPase
title_full Chemical genetic identification of GAK substrates reveals its role in regulating Na(+)/K(+)-ATPase
title_fullStr Chemical genetic identification of GAK substrates reveals its role in regulating Na(+)/K(+)-ATPase
title_full_unstemmed Chemical genetic identification of GAK substrates reveals its role in regulating Na(+)/K(+)-ATPase
title_short Chemical genetic identification of GAK substrates reveals its role in regulating Na(+)/K(+)-ATPase
title_sort chemical genetic identification of gak substrates reveals its role in regulating na(+)/k(+)-atpase
topic Research Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6312924/
https://www.ncbi.nlm.nih.gov/pubmed/30623173
http://dx.doi.org/10.26508/lsa.201800118
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