Hydrogen peroxide-dependent oxidation of ERK2 within its D-recruitment site alters its substrate selection

Anthony E Postiglione; Laquaundra L Adams; Ese S Ekhator; Anuoluwapo E Odelade; Supriya Patwardhan; Meenal Chaudhari; Avery S Pardue; Anjali Kumari; William A LeFever; Olivia P Tornow; Tamer S Kaoud; Johnathan Neiswinger; Jun Seop Jeong; Derek Parsonage; Kimberly J Nelson; Dukka B Kc; Cristina M Furdui; Heng Zhu; Andrew J Wommack; Kevin N Dalby; Ming Dong; Leslie B Poole; Jeremiah D Keyes; Robert H Newman

doi:10.1016/j.isci.2023.107817

Hydrogen peroxide-dependent oxidation of ERK2 within its D-recruitment site alters its substrate selection

iScience. 2023 Sep 2;26(10):107817. doi: 10.1016/j.isci.2023.107817. eCollection 2023 Oct 20.

Authors

Anthony E Postiglione^{1

2}, Laquaundra L Adams¹, Ese S Ekhator¹, Anuoluwapo E Odelade¹, Supriya Patwardhan¹, Meenal Chaudhari^{1

3

4}, Avery S Pardue¹, Anjali Kumari¹, William A LeFever^{5

6}, Olivia P Tornow⁵, Tamer S Kaoud⁷, Johnathan Neiswinger^{8

9}, Jun Seop Jeong¹, Derek Parsonage¹⁰, Kimberly J Nelson¹⁰, Dukka B Kc¹¹, Cristina M Furdui¹², Heng Zhu⁸, Andrew J Wommack⁵, Kevin N Dalby⁷, Ming Dong^{13

14}, Leslie B Poole¹⁰, Jeremiah D Keyes^{10

15

16}, Robert H Newman¹

Affiliations

¹ Department of Biology, North Carolina A&T State University, Greensboro, NC 27411, USA.
² Department of Biology, Wake Forest University, Winston-Salem, NC 27101, USA.
³ Department of Computational Data Science and Engineering, North Carolina A&T State University, Greensboro, NC 27411, USA.
⁴ Department of Mathematics and Computer Science, University of Virginia at Wise, Wise, VA 24293, USA.
⁵ Department of Chemistry, High Point University, High Point, NC 27268, USA.
⁶ Department of Chemistry, Purdue University, West Lafayette, IN 47907, USA.
⁷ Division of Chemical Biology and Medicinal Chemistry, The University of Texas at Austin, Austin, TX 78712, USA.
⁸ Department of Pharmacology and Molecular Sciences, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
⁹ Department of Biology, Belhaven University, Jackson, MS 39202, USA.
¹⁰ Department of Biochemistry, Wake Forest University School of Medicine, Winston-Salem, NC 27157, USA.
¹¹ Department of Computer Science, Michigan Technological University, Houghton, MI 49931, USA.
¹² Department of Internal Medicine, Section on Molecular Medicine, Wake Forest University School of Medicine, Winston-Salem, NC 27157, USA.
¹³ Department of Chemistry, North Carolina A&T State University, Greensboro, NC 27411, USA.
¹⁴ Department of Chemistry and Biochemistry, University of North Carolina Wilmington, Wilmington, NC 28403, USA.
¹⁵ Department of Biology, Penn State University Behrend, Erie, PA 16563, USA.
¹⁶ Magee-Womens Research Institute, Pittsburgh, PA 15213, USA.

Abstract

Extracellular signal-regulated kinases 1 and 2 (ERK1/2) are dysregulated in many pervasive diseases. Recently, we discovered that ERK1/2 is oxidized by signal-generated hydrogen peroxide in various cell types. Since the putative sites of oxidation lie within or near ERK1/2's ligand-binding surfaces, we investigated how oxidation of ERK2 regulates interactions with the model substrates Sub-D and Sub-F. These studies revealed that ERK2 undergoes sulfenylation at C159 on its D-recruitment site surface and that this modification modulates ERK2 activity differentially between substrates. Integrated biochemical, computational, and mutational analyses suggest a plausible mechanism for peroxide-dependent changes in ERK2-substrate interactions. Interestingly, oxidation decreased ERK2's affinity for some D-site ligands while increasing its affinity for others. Finally, oxidation by signal-generated peroxide enhanced ERK1/2's ability to phosphorylate ribosomal S6 kinase A1 (RSK1) in HeLa cells. Together, these studies lay the foundation for examining crosstalk between redox- and phosphorylation-dependent signaling at the level of kinase-substrate selection.

Keywords: Biochemistry; Biological sciences; Cell biology; Molecular biology.

Grants and funding

R35 GM135179/GM/NIGMS NIH HHS/United States