Novak Leonore, Petrosino Maria, Pasquo Alessandra, Chaikuad Apirat, Chiaraluce Roberta, Knapp Stefan, Consalvi Valerio
Dipartimento di Scienze Biochimiche "A. Rossi Fanelli", Sapienza University of Rome, 00185 Rome, Italy.
Chair of Pharmacology, Section of Medicine, University of Fribourg, CH-1700 Fribourg, Switzerland.
Cancers (Basel). 2023 May 26;15(11):2938. doi: 10.3390/cancers15112938.
The extracellular-signal-regulated kinase 2 (ERK2), a mitogen-activated protein kinase (MAPK) located downstream of the Ras-Raf-MEK-ERK signal transduction cascade, is involved in the regulation of a large variety of cellular processes. The ERK2, activated by phosphorylation, is the principal effector of a central signaling cascade that converts extracellular stimuli into cells. Deregulation of the ERK2 signaling pathway is related to many human diseases, including cancer. This study reports a comprehensive biophysical analysis of structural, function, and stability data of pure, recombinant human non-phosphorylated (NP-) and phosphorylated (P-) ERK2 wild-type and missense variants in the common docking site (CD-site) found in cancer tissues. Because the CD-site is involved in interaction with protein substrates and regulators, a biophysical characterization of missense variants adds information about the impact of point mutations on the ERK2 structure-function relationship. Most of the P-ERK2 variants in the CD-site display a reduced catalytic efficiency, and for the P-ERK2 D321E, D321N, D321V and E322K, changes in thermodynamic stability are observed. The thermal stability of NP-ERK2 and P-ERK2 D321E, D321G, and E322K is decreased with respect to the wild-type. In general, a single residue mutation in the CD-site may lead to structural local changes that reflects in alterations in the global ERK2 stability and catalysis.
细胞外信号调节激酶2（ERK2）是一种丝裂原活化蛋白激酶（MAPK），位于Ras-Raf-MEK-ERK信号转导级联的下游，参与多种细胞过程的调控。ERK2通过磷酸化激活，是将细胞外刺激转化为细胞内信号的核心信号级联的主要效应器。ERK2信号通路的失调与许多人类疾病有关，包括癌症。本研究报告了对癌症组织中常见对接位点（CD位点）的纯合重组人非磷酸化（NP-）和磷酸化（P-）ERK2野生型和错义变体的结构、功能和稳定性数据的全面生物物理分析。由于CD位点参与与蛋白质底物和调节剂的相互作用，错义变体的生物物理特征增加了关于点突变对ERK2结构-功能关系影响的信息。CD位点中的大多数P-ERK2变体显示出催化效率降低，对于P-ERK2 D321E、D321N、D321V和E322K，观察到热力学稳定性的变化。与野生型相比，NP-ERK2和P-ERK2 D321E、D321G和E322K的热稳定性降低。一般来说，CD位点中的单个残基突变可能导致结构局部变化，这反映在ERK2整体稳定性和催化作用的改变上。
