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Structural insights and biophysical characterization of p90RSK2:ERK2 complex
Evan Kobori1, Hoang Nguyen1, Jian Wu2
1Department of Chemistry and Biochemistry, University of California, San Diego, CA, U.S.A.
The Biochemical Journal
|July 30, 2025
Summary
Studying the full-length RSK2:ERK2 kinase complex reveals ERK2 is more protected. Complementary techniques like HDXMS and cryoEM provide insights into these challenging kinase structures.
Area of Science:
- Biochemistry and structural biology
- Protein dynamics and complex formation
Background:
- Kinase function relies on interplay between ordered domains and flexible intrinsically disordered regions (IDRs).
- Full-length kinase characterization is challenging due to IDRs, yet crucial for understanding regulation.
- The p90 ribosomal S6 kinase (RSK) family possesses two functional kinase domains (NTK and CTK).
Purpose of the Study:
- To investigate the structure and biophysical characteristics of the full-length RSK2:ERK2 complex.
- To understand the interplay between RSK2's kinase domains and its activator, ERK2.
- To demonstrate the utility of complementary techniques for studying challenging kinase complexes.
Main Methods:
- Hydrogen-deuterium exchange mass spectrometry (HDXMS) for solvent accessibility.
- Cryo-electron microscopy (cryoEM) for structural determination.
- AlphaFold modeling for computational structure prediction.
Main Results:
- ERK2 exhibited greater solvent protection compared to the RSK2 NTK/CTK domains.
- The RSK2 NTK N-lobe showed high deuterium uptake, indicating dynamic conformational flexibility.
- CryoEM and AlphaFold modeling suggested a compact RSK2:ERK2 complex with a potential NTK-ERK2 interface.
Conclusions:
- Multiple complementary biophysical techniques offer valuable insights into complex kinase structures.
- The RSK2:ERK2 complex exhibits specific structural features and domain dynamics.
- Understanding these dynamics is key to RSK kinase function and regulation.
