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Mutation in the Common Docking Domain Affects MAP Kinase ERK2 Catalysis and Stability

Leonore Novak1, Maria Petrosino2, Alessandra Pasquo3

  • 1Dipartimento di Scienze Biochimiche "A. Rossi Fanelli", Sapienza University of Rome, 00185 Rome, Italy.

Cancers
|June 10, 2023
PubMed

Insights

Mutations in the ERK2 docking site impact its function and stability. This study analyzes how these changes affect cancer-related signaling pathways, revealing potential therapeutic targets.

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Cellular Signaling

Background:

  • Extracellular-signal-regulated kinase 2 (ERK2) is a key mitogen-activated protein kinase (MAPK) in the Ras-Raf-MEK-ERK pathway.
  • ERK2 signaling is crucial for cellular processes and its deregulation is linked to human diseases, notably cancer.
  • The common docking site (CD-site) of ERK2 is vital for substrate and regulator interactions.

Purpose of the Study:

  • To conduct a comprehensive biophysical analysis of wild-type and variant human ERK2 (non-phosphorylated and phosphorylated).
  • To investigate the impact of missense mutations in the ERK2 CD-site on its structure, function, and stability.
  • To understand how point mutations affect the ERK2 structure-function relationship in the context of cancer.

Main Methods:

  • Purification of recombinant human non-phosphorylated (NP-) and phosphorylated (P-) ERK2 wild-type and missense variants.
  • Comprehensive biophysical analysis including structural, functional, and stability assays.
  • Characterization of variants located in the common docking site (CD-site).

Main Results:

  • Most P-ERK2 variants in the CD-site exhibited reduced catalytic efficiency.
  • Specific variants (P-ERK2 D321E, D321N, D321V, E322K) showed altered thermodynamic stability.
  • Thermal stability was decreased in NP-ERK2 and P-ERK2 variants (D321E, D321G, E322K) compared to wild-type.

Conclusions:

  • Single residue mutations in the ERK2 CD-site can induce local structural changes.
  • These structural alterations lead to changes in global ERK2 stability and catalytic activity.
  • Understanding these mutations' effects is crucial for deciphering ERK2's role in disease and developing targeted therapies.
Keywords:
ERK2MAP kinaseMAPK1cancer mutations and protein structure alterationsmutationprotein stabilitysingle nucleotide variant

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