Activation and inhibition of the C-terminal kinase domain of p90 ribosomal S6 kinases.

Marlene Uglebjerg Fruergaard, Christine Juul Fælled Nielsen, Cecilia Rosada Kjeldsen, Lars Iversen, Jacob Lauwring Andersen, Poul Nissen

Life science alliance 2023 May

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The p90 ribosomal S6 kinases (RSKs) contain two distinct catalytic kinase domains, the N-terminal and C-terminal kinase domains (NTKD and CTKD, respectively). The activation of CTKD is regulated by phosphorylation by extracellular signal-regulated kinase (ERK1/2) and an autoinhibitory αL helix. Through a mutational series in vitro of the RSK CTKDs, we found a complex mechanism lifting autoinhibition that led us to design constitutively active RSK CTKDs. These are based on a phosphomimetic mutation and a C-terminal truncation (e.g., RSK2 T577E D694*) where a high activity in absence of ERK phosphorylation is obtained. Using these constructs, we characterize IC50 values of ATP-competitive inhibitors and provide a setup for determining specificity constants (kinact/Ki) of covalent CTKD inhibitors. © 2023 Fruergaard et al.

Citation

Marlene Uglebjerg Fruergaard, Christine Juul Fælled Nielsen, Cecilia Rosada Kjeldsen, Lars Iversen, Jacob Lauwring Andersen, Poul Nissen. Activation and inhibition of the C-terminal kinase domain of p90 ribosomal S6 kinases. Life science alliance. 2023 May;6(5)