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Isoform-selective activity-based profiling of ERK signaling
Extracellular signal-regulated kinases (ERKs) mediate downstream signaling of RAS-RAF-MEK as key regulators of the mitogen-activated protein kinase (MAPK) pathway. Activation of ERK signaling is a hallmark of cancer and upstream MAPK proteins have been extensively pursued as drug targets for cancer...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Royal Society of Chemistry
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5909473/ https://www.ncbi.nlm.nih.gov/pubmed/29732117 http://dx.doi.org/10.1039/c8sc00043c |
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author | Shin, Myungsun Franks, Caroline E. Hsu, Ku-Lung |
author_facet | Shin, Myungsun Franks, Caroline E. Hsu, Ku-Lung |
author_sort | Shin, Myungsun |
collection | PubMed |
description | Extracellular signal-regulated kinases (ERKs) mediate downstream signaling of RAS-RAF-MEK as key regulators of the mitogen-activated protein kinase (MAPK) pathway. Activation of ERK signaling is a hallmark of cancer and upstream MAPK proteins have been extensively pursued as drug targets for cancer therapies. However, the rapid rise of resistance to clinical RAF and MEK inhibitors has prompted interest in targeting ERK (ERK1 and ERK2 isoforms) directly for cancer therapy. Current methods for evaluating activity of inhibitors against ERK isoforms are based primarily on analysis of recombinant proteins. Strategies to directly and independently profile native ERK1 and ERK2 activity would greatly complement current cell biological tools used to probe and target ERK function. Here, we present a quantitative chemoproteomic strategy that utilizes active-site directed probes to directly quantify native ERK activity in an isoform-specific fashion. We exploit a single isoleucine/leucine difference in ERK substrate binding sites to enable activity-based profiling of ERK1 versus ERK2 across a variety of cell types, tissues, and species. We used our chemoproteomic strategy to determine potency and selectivity of academic (VX-11e) and clinical (Ulixertinib) ERK inhibitors. Correlation of potency estimates by chemoproteomics with anti-proliferative activity of VX-11e and Ulixertinib revealed that >90% inactivation of both native ERK1 and ERK2 is needed to mediate cellular activity of inhibitors. Our findings introduce one of the first assays capable of independent evaluation of native ERK1 and ERK2 activity to advance drug discovery of oncogenic MAPK pathways. |
format | Online Article Text |
id | pubmed-5909473 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | Royal Society of Chemistry |
record_format | MEDLINE/PubMed |
spelling | pubmed-59094732018-05-04 Isoform-selective activity-based profiling of ERK signaling Shin, Myungsun Franks, Caroline E. Hsu, Ku-Lung Chem Sci Chemistry Extracellular signal-regulated kinases (ERKs) mediate downstream signaling of RAS-RAF-MEK as key regulators of the mitogen-activated protein kinase (MAPK) pathway. Activation of ERK signaling is a hallmark of cancer and upstream MAPK proteins have been extensively pursued as drug targets for cancer therapies. However, the rapid rise of resistance to clinical RAF and MEK inhibitors has prompted interest in targeting ERK (ERK1 and ERK2 isoforms) directly for cancer therapy. Current methods for evaluating activity of inhibitors against ERK isoforms are based primarily on analysis of recombinant proteins. Strategies to directly and independently profile native ERK1 and ERK2 activity would greatly complement current cell biological tools used to probe and target ERK function. Here, we present a quantitative chemoproteomic strategy that utilizes active-site directed probes to directly quantify native ERK activity in an isoform-specific fashion. We exploit a single isoleucine/leucine difference in ERK substrate binding sites to enable activity-based profiling of ERK1 versus ERK2 across a variety of cell types, tissues, and species. We used our chemoproteomic strategy to determine potency and selectivity of academic (VX-11e) and clinical (Ulixertinib) ERK inhibitors. Correlation of potency estimates by chemoproteomics with anti-proliferative activity of VX-11e and Ulixertinib revealed that >90% inactivation of both native ERK1 and ERK2 is needed to mediate cellular activity of inhibitors. Our findings introduce one of the first assays capable of independent evaluation of native ERK1 and ERK2 activity to advance drug discovery of oncogenic MAPK pathways. Royal Society of Chemistry 2018-02-06 /pmc/articles/PMC5909473/ /pubmed/29732117 http://dx.doi.org/10.1039/c8sc00043c Text en This journal is © The Royal Society of Chemistry 2018 http://creativecommons.org/licenses/by/3.0/ This article is freely available. This article is licensed under a Creative Commons Attribution 3.0 Unported Licence (CC BY 3.0) |
spellingShingle | Chemistry Shin, Myungsun Franks, Caroline E. Hsu, Ku-Lung Isoform-selective activity-based profiling of ERK signaling |
title | Isoform-selective activity-based profiling of ERK signaling
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title_full | Isoform-selective activity-based profiling of ERK signaling
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title_fullStr | Isoform-selective activity-based profiling of ERK signaling
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title_full_unstemmed | Isoform-selective activity-based profiling of ERK signaling
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title_short | Isoform-selective activity-based profiling of ERK signaling
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title_sort | isoform-selective activity-based profiling of erk signaling |
topic | Chemistry |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5909473/ https://www.ncbi.nlm.nih.gov/pubmed/29732117 http://dx.doi.org/10.1039/c8sc00043c |
work_keys_str_mv | AT shinmyungsun isoformselectiveactivitybasedprofilingoferksignaling AT frankscarolinee isoformselectiveactivitybasedprofilingoferksignaling AT hsukulung isoformselectiveactivitybasedprofilingoferksignaling |