@article{7c39f529592946c4b42b84b1e085d693,
title = "Functional divergence caused by mutations in an energetic hotspot in ERK2",
abstract = "The most frequent extracellular signal-regulated kinase 2 (ERK2) mutation occurring in cancers is E322K (E-K). ERK2 E-K reverses a buried charge in the ERK2 common docking (CD) site a region that binds activators inhibitors and substrates. Little is known about the cellular consequences associated with this mutation other than apparent increases in tumor resistance to pathway inhibitors. ERK2 E-K like the mutation of the preceding aspartate (ERK2 D321N [D-N]) known as the sevenmaker mutation causes increased activity in cells and evades inactivation by dual-specificity phosphatases. As opposed to findings in cancer cells in developmental assays in Drosophila only ERK2 D-N displays a significant gain of function revealing mutation-specific phenotypes. The crystal structure of ERK2 D-N is indistinguishable from that of wild-type protein yet this mutant displays increased thermal stability. In contrast the crystal structure of ERK2 E-K reveals profound structural changes including disorder in the CD site and exposure of the activation loop phosphorylation sites which likely account for the decreased thermal stability of the protein. These contiguous mutations in the CD site of ERK2 are both required for docking interactions but lead to unpredictably different functional outcomes. Our results suggest that the CD site is in an energetically strained configuration and this helps drive conformational changes at distal sites on ERK2 during docking interactions.",
keywords = "ERK CD site, Kinase, Mutation, Stability",
author = "Taylor, {Clinton A.} and Cormier, {Kevin W.} and Keenan, {Shannon E.} and Svetlana Earnest and Steve Stippec and Chonlarat Wichaidit and Juang, {Yu Chi} and Junmei Wang and Shvartsman, {Stanislav Y.} and Goldsmith, {Elizabeth J.} and Cobb, {Melanie H.}",
note = "Funding Information: ACKNOWLEDGMENTS. We thank Dominika Borek, Diana Tomchick, and Zhe Chen (Department of Biophysics, UT Southwestern) for advice and assistance with crystallization, data collection, and data processing; Chad Brautigam and Shih-Chia Tso (Department of Biophysics, UT Southwestern) for assistance with dynamic light scattering and microscale thermophoresis; Radha Akella (Department of Biophysics, UT Southwestern) for assistance with thermal stability assays; Dean Smith and Ben Weaver (Department of Pharmacology, UT Southwestern) for thoughtful comments on the manuscript; Michael Reese (Department of Pharmacology, UT Southwestern) for suggestions about plotting B-factor data; Aroon Karra, Wen-Huang Ko, Jihan Osborne, and other current and former members of the M.H.C. laboratory for their advice and assistance with specific aspects of this project; and Dionne Ware for administrative assistance. Results shown in this report are derived from work performed at the Argonne National Laboratory, Structural Biology Center at the Advanced Photon Source. The Argonne National Laboratory is operated by the University of Chicago Argonne, LLC, for the US Department of Energy, Office of Biological and Environmental Research, under contract DE-AC02-06CH11357. This project was supported by Welch Foundation Grants I1243 (to M.H.C.) and I1128 (to E.J.G.) and by NIH Grants R37DK34128 (to M.H.C.) and R01GM086537 (to S.Y.S.). C.A.T. was supported in the late stages of this work by Cancer Prevention and Research Institute of Texas Training Grant RP160157. Additional support was provided by National Cancer Institute Grant P30CA142543 to the Harold C. Simmons Comprehensive Cancer Center for core facilities. Publisher Copyright: {\textcopyright} 2019 National Academy of Sciences. All rights reserved.",
year = "2019",
month = jul,
day = "30",
doi = "10.1073/pnas.1905015116",
language = "English (US)",
volume = "116",
pages = "15514--15523",
journal = "Proceedings of the National Academy of Sciences of the United States of America",
issn = "0027-8424",
publisher = "National Academy of Sciences",
number = "31",
}