Fluorophore labeled kinase detects ligands that bind within the MAPK insert of p38α kinase.

The vast majority of small molecules known to modulate kinase activity, target the highly conserved ATP-pocket. Consequently, such ligands are often less specific and in case of inhibitors, this leads to the inhibition of multiple kinases. Thus, selective modulation of kinase function remains a majo...

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Autores principales:	Matthäus Getlik, Jeffrey R Simard, Martin Termathe, Christian Grütter, Matthias Rabiller, Willem A L van Otterlo, Daniel Rauh
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2012
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/87ccca3039fb4ca1a25928d702a9a1d7
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spelling	oai:doaj.org-article:87ccca3039fb4ca1a25928d702a9a1d72021-11-18T07:13:34ZFluorophore labeled kinase detects ligands that bind within the MAPK insert of p38α kinase.1932-620310.1371/journal.pone.0039713https://doaj.org/article/87ccca3039fb4ca1a25928d702a9a1d72012-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/22768308/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203The vast majority of small molecules known to modulate kinase activity, target the highly conserved ATP-pocket. Consequently, such ligands are often less specific and in case of inhibitors, this leads to the inhibition of multiple kinases. Thus, selective modulation of kinase function remains a major hurdle. One of the next great challenges in kinase research is the identification of ligands which bind to less conserved sites and target the non-catalytic functions of protein kinases. However, approaches that allow for the unambiguous identification of molecules that bind to these less conserved sites are few in number. We have previously reported the use of fluorescent labels in kinases (FLiK) to develop direct kinase binding assays that exclusively detect ligands which stabilize inactive (DFG-out) kinase conformations. Here, we present the successful application of the FLiK approach to develop a high-throughput binding assay capable of directly monitoring ligand binding to a remote site within the MAPK insert of p38α mitogen-activated protein kinase (MAPK). Guided by the crystal structure of an initially identified hit molecule in complex with p38α, we developed a tight binding ligand which may serve as an ideal starting point for further investigations of the biological function of the MAPK insert in regulating the p38α signaling pathway.Matthäus GetlikJeffrey R SimardMartin TermatheChristian GrütterMatthias RabillerWillem A L van OtterloDaniel RauhPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 7, Iss 7, p e39713 (2012)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Matthäus Getlik Jeffrey R Simard Martin Termathe Christian Grütter Matthias Rabiller Willem A L van Otterlo Daniel Rauh Fluorophore labeled kinase detects ligands that bind within the MAPK insert of p38α kinase.
description	The vast majority of small molecules known to modulate kinase activity, target the highly conserved ATP-pocket. Consequently, such ligands are often less specific and in case of inhibitors, this leads to the inhibition of multiple kinases. Thus, selective modulation of kinase function remains a major hurdle. One of the next great challenges in kinase research is the identification of ligands which bind to less conserved sites and target the non-catalytic functions of protein kinases. However, approaches that allow for the unambiguous identification of molecules that bind to these less conserved sites are few in number. We have previously reported the use of fluorescent labels in kinases (FLiK) to develop direct kinase binding assays that exclusively detect ligands which stabilize inactive (DFG-out) kinase conformations. Here, we present the successful application of the FLiK approach to develop a high-throughput binding assay capable of directly monitoring ligand binding to a remote site within the MAPK insert of p38α mitogen-activated protein kinase (MAPK). Guided by the crystal structure of an initially identified hit molecule in complex with p38α, we developed a tight binding ligand which may serve as an ideal starting point for further investigations of the biological function of the MAPK insert in regulating the p38α signaling pathway.
format	article
author	Matthäus Getlik Jeffrey R Simard Martin Termathe Christian Grütter Matthias Rabiller Willem A L van Otterlo Daniel Rauh
author_facet	Matthäus Getlik Jeffrey R Simard Martin Termathe Christian Grütter Matthias Rabiller Willem A L van Otterlo Daniel Rauh
author_sort	Matthäus Getlik
title	Fluorophore labeled kinase detects ligands that bind within the MAPK insert of p38α kinase.
title_short	Fluorophore labeled kinase detects ligands that bind within the MAPK insert of p38α kinase.
title_full	Fluorophore labeled kinase detects ligands that bind within the MAPK insert of p38α kinase.
title_fullStr	Fluorophore labeled kinase detects ligands that bind within the MAPK insert of p38α kinase.
title_full_unstemmed	Fluorophore labeled kinase detects ligands that bind within the MAPK insert of p38α kinase.
title_sort	fluorophore labeled kinase detects ligands that bind within the mapk insert of p38α kinase.
publisher	Public Library of Science (PLoS)
publishDate	2012
url	https://doaj.org/article/87ccca3039fb4ca1a25928d702a9a1d7
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Fluorophore labeled kinase detects ligands that bind within the MAPK insert of p38α kinase.

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