@article{24e3425a91434ee5aa80547015bbaed4,
title = "Regulation of the EphA2 receptor intracellular region by phosphomimetic negative charges in the kinase-SAM linker",
abstract = "Eph receptor tyrosine kinases play a key role in cell-cell communication. Lack of structural information on the entire multi-domain intracellular region of any Eph receptor has hindered understanding of their signaling mechanisms. Here, we use integrative structural biology to investigate the structure and dynamics of the EphA2 intracellular region. EphA2 promotes cancer malignancy through a poorly understood non-canonical form of signaling involving serine/threonine phosphorylation of the linker connecting its kinase and SAM domains. We show that accumulation of multiple linker negative charges, mimicking phosphorylation, induces cooperative changes in the EphA2 intracellular region from more closed to more extended conformations and perturbs the EphA2 juxtamembrane segment and kinase domain. In cells, linker negative charges promote EphA2 oligomerization. We also identify multiple kinases catalyzing linker phosphorylation. Our findings suggest multiple effects of linker phosphorylation on EphA2 signaling and imply that coordination of different kinases is necessary to promote EphA2 non-canonical signaling.",
author = "Lechtenberg, {Bernhard C.} and Gehring, {Marina P.} and Light, {Taylor P.} and Horne, {Christopher R.} and Matsumoto, {Mike W.} and Kalina Hristova and Pasquale, {Elena B.}",
note = "Funding Information: The authors thank P. Hornbeck (PhosphoSitePlus) for providing original data on phosphosites in individual peptides from the PhosphoSite database, Alejandro Conde-Perez for generating the MEL-JUSO-EphA2 stable cell line, B. Emerling for providing the SW626 cell line, P. Itkin-Ansari for the BxPC3 cell line, F. Levine for the PANC1 pancreatic cancer cell line, K. Vuori for the SKOV3 and MDA-MB-468 cell lines, J. Smith for the MDA-MB-231 cell line, R. Maki for the BT549 cell line, W. Stallcup for the U87, T98 and U251-MG cell lines, the staff of the BioCAT Advanced SAXS Training Course, in particular W. Shang, for support with initial SAXS data collection and analysis, staff of the Australian Synchrotron SAXS/WAXS beamline (part of ANSTO) for technical assistance, J. Murphy (WEHI) for help with SAXS data collection and analysis, A. Bobkov for collecting and analyzing AUC data, S. Silletti, UCSD Biomolecular and Proteomics Mass Spectrometry (BPSM) Facility, for HDX-MS data collection and analysis and Ryan Lumpkin for help with DECA. This work was supported by NIH grants GM131374 and AG062617 and institutional funds to EBP, NIH grant GM068619 to KH, and National Cancer Institute Cancer Center Support grant P30 CA030199, which supported SBP Core Facilities and funds for a pilot project. The UCSD BPSM Facility, which carried out the HDX experiments, is supported by the NIH shared instrumentation grant number S10 OD016234 (Synapt-HDX-MS). This project also used resources of the Advanced Photon Source, a U.S. Department of Energy (DOE) Office of Science User Facility operated for the DOE Office of Science by Argonne National Laboratory under Contract No. DE-AC02-06CH11357. The work for this project was supported by grant 9 P41 GM103622 from the National Institute of General Medical Sciences of the National Institutes of Health. Use of the BioCAT Pilatus 3 1 M detector was provided by grant 1S10OD018090-01 from NIGMS. The content is solely the responsibility of the authors and does not necessarily reflect the official views of the National Institute of General Medical Sciences or the National Institutes of Health. Publisher Copyright: {\textcopyright} 2021, The Author(s).",
year = "2021",
month = dec,
doi = "10.1038/s41467-021-27343-z",
language = "English (US)",
volume = "12",
journal = "Nature Communications",
issn = "2041-1723",
publisher = "Nature Publishing Group",
number = "1",
}