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

doi:10.26508/lsa.202201425

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

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaper › Journal article › Research › peer-review

Abstract

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.

Original language	English
Article number	e202201425
Journal	Life Science Alliance
Volume	6
Issue	5
ISSN	2575-1077
DOIs	https://doi.org/10.26508/lsa.202201425
Publication status	Published - May 2023

Access to Document

10.26508/lsa.202201425

Cite this

@article{582032b1d35a45708529c30d770b2e28,

title = "Activation and inhibition of the C-terminal kinase domain of p90 ribosomal S6 kinases",

abstract = "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.",

author = "Fruergaard, {Marlene Uglebjerg} and Nielsen, {Christine Juul F{\ae}lled} and Kjeldsen, {Cecilia Rosada} and Lars Iversen and Andersen, {Jacob Lauwring} and Poul Nissen",

note = "Publisher Copyright: {\textcopyright} 2023 Fruergaard et al.",

year = "2023",

month = may,

doi = "10.26508/lsa.202201425",

language = "English",

volume = "6",

journal = "Life Science Alliance",

issn = "2575-1077",

publisher = "Rockefeller University Press",

number = "5",

}

Activation and inhibition of the C-terminal kinase domain of p90 ribosomal S6 kinases. / Fruergaard, Marlene Uglebjerg ; Nielsen, Christine Juul Fælled; Kjeldsen, Cecilia Rosada et al.
In: Life Science Alliance, Vol. 6, No. 5, e202201425, 05.2023.

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaper › Journal article › Research › peer-review

TY - JOUR

T1 - Activation and inhibition of the C-terminal kinase domain of p90 ribosomal S6 kinases

AU - Fruergaard, Marlene Uglebjerg

AU - Nielsen, Christine Juul Fælled

AU - Kjeldsen, Cecilia Rosada

AU - Iversen, Lars

AU - Andersen, Jacob Lauwring

AU - Nissen, Poul

PY - 2023/5

Y1 - 2023/5

N2 - 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.

AB - 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.

U2 - 10.26508/lsa.202201425

DO - 10.26508/lsa.202201425

M3 - Journal article

C2 - 36806093

AN - SCOPUS:85148396469

SN - 2575-1077

VL - 6

JO - Life Science Alliance

JF - Life Science Alliance

IS - 5

M1 - e202201425

ER -

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

Abstract

Access to Document

Fingerprint

Cite this