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Target-Based Discovery of an Inhibitor of the Regulatory Phosphatase PPP1R15B

Protein phosphorylation is a prevalent and ubiquitous mechanism of regulation. Kinases are popular drug targets, but identifying selective phosphatase inhibitors has been challenging. Here, we used surface plasmon resonance to design a method to enable target-based discovery of selective serine/thre...

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Autores principales: Krzyzosiak, Agnieszka, Sigurdardottir, Anna, Luh, Laura, Carrara, Marta, Das, Indrajit, Schneider, Kim, Bertolotti, Anne
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Cell Press 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6108835/
https://www.ncbi.nlm.nih.gov/pubmed/30057111
http://dx.doi.org/10.1016/j.cell.2018.06.030
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author Krzyzosiak, Agnieszka
Sigurdardottir, Anna
Luh, Laura
Carrara, Marta
Das, Indrajit
Schneider, Kim
Bertolotti, Anne
author_facet Krzyzosiak, Agnieszka
Sigurdardottir, Anna
Luh, Laura
Carrara, Marta
Das, Indrajit
Schneider, Kim
Bertolotti, Anne
author_sort Krzyzosiak, Agnieszka
collection PubMed
description Protein phosphorylation is a prevalent and ubiquitous mechanism of regulation. Kinases are popular drug targets, but identifying selective phosphatase inhibitors has been challenging. Here, we used surface plasmon resonance to design a method to enable target-based discovery of selective serine/threonine phosphatase inhibitors. The method targeted a regulatory subunit of protein phosphatase 1, PPP1R15B (R15B), a negative regulator of proteostasis. This yielded Raphin1, a selective inhibitor of R15B. In cells, Raphin1 caused a rapid and transient accumulation of its phosphorylated substrate, resulting in a transient attenuation of protein synthesis. In vitro, Raphin1 inhibits the recombinant R15B-PP1c holoenzyme, but not the closely related R15A-PP1c, by interfering with substrate recruitment. Raphin1 was orally bioavailable, crossed the blood-brain barrier, and demonstrated efficacy in a mouse model of Huntington’s disease. This identifies R15B as a druggable target and provides a platform for target-based discovery of inhibitors of serine/threonine phosphatases.
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spelling pubmed-61088352018-08-27 Target-Based Discovery of an Inhibitor of the Regulatory Phosphatase PPP1R15B Krzyzosiak, Agnieszka Sigurdardottir, Anna Luh, Laura Carrara, Marta Das, Indrajit Schneider, Kim Bertolotti, Anne Cell Article Protein phosphorylation is a prevalent and ubiquitous mechanism of regulation. Kinases are popular drug targets, but identifying selective phosphatase inhibitors has been challenging. Here, we used surface plasmon resonance to design a method to enable target-based discovery of selective serine/threonine phosphatase inhibitors. The method targeted a regulatory subunit of protein phosphatase 1, PPP1R15B (R15B), a negative regulator of proteostasis. This yielded Raphin1, a selective inhibitor of R15B. In cells, Raphin1 caused a rapid and transient accumulation of its phosphorylated substrate, resulting in a transient attenuation of protein synthesis. In vitro, Raphin1 inhibits the recombinant R15B-PP1c holoenzyme, but not the closely related R15A-PP1c, by interfering with substrate recruitment. Raphin1 was orally bioavailable, crossed the blood-brain barrier, and demonstrated efficacy in a mouse model of Huntington’s disease. This identifies R15B as a druggable target and provides a platform for target-based discovery of inhibitors of serine/threonine phosphatases. Cell Press 2018-08-23 /pmc/articles/PMC6108835/ /pubmed/30057111 http://dx.doi.org/10.1016/j.cell.2018.06.030 Text en © 2018 MRC Laboratory of Molecular Biology http://creativecommons.org/licenses/by/4.0/ This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Krzyzosiak, Agnieszka
Sigurdardottir, Anna
Luh, Laura
Carrara, Marta
Das, Indrajit
Schneider, Kim
Bertolotti, Anne
Target-Based Discovery of an Inhibitor of the Regulatory Phosphatase PPP1R15B
title Target-Based Discovery of an Inhibitor of the Regulatory Phosphatase PPP1R15B
title_full Target-Based Discovery of an Inhibitor of the Regulatory Phosphatase PPP1R15B
title_fullStr Target-Based Discovery of an Inhibitor of the Regulatory Phosphatase PPP1R15B
title_full_unstemmed Target-Based Discovery of an Inhibitor of the Regulatory Phosphatase PPP1R15B
title_short Target-Based Discovery of an Inhibitor of the Regulatory Phosphatase PPP1R15B
title_sort target-based discovery of an inhibitor of the regulatory phosphatase ppp1r15b
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6108835/
https://www.ncbi.nlm.nih.gov/pubmed/30057111
http://dx.doi.org/10.1016/j.cell.2018.06.030
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