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Stabilization of Protein-Protein Interactions between CaMKK2 and 14-3-3 by Fusicoccins

SYSNO ASEP	0535685
Document Type	J - Journal Article
R&D Document Type	Journal Article
Subsidiary J	Článek ve WOS
Title	Stabilization of Protein-Protein Interactions between CaMKK2 and 14-3-3 by Fusicoccins
Author(s)	Santo Lentini, D. (CZ) Petrvalská, Olivia (FGU-C)_{RID, ORCID, SAI} Obšilová, Veronika (FGU-C)_{RID, ORCID, SAI} Ottmann, Ch. (NL) Obšil, Tomáš (FGU-C)_{RID, ORCID}
Source Title	ACS Chemical Biology. - : American Chemical Society - ISSN 1554-8929 Roč. 15, č. 11 (2020), s. 3060-3071
Number of pages	12 s.
Language	eng - English
Country	US - United States
Keywords	CaMKK2 ; 14-3-3 ; protein-protein interaction ; fusicoccin ; structure stabilization
Subject RIV	CE - Biochemistry
OECD category	Biochemistry and molecular biology
R&D Projects	GA19-00121S GA ČR - Czech Science Foundation (CSF)
Research Infrastructure	CIISB II - 90127 - Masarykova univerzita
Method of publishing	Limited access
Institutional support	FGU-C - RVO:67985823
UT WOS	000592981100024
EID SCOPUS	85096508476
DOI	10.1021/acschembio.0c00821
Annotation	Ca2+/calmodulin-dependent protein kinase kinase 2 (CaMKK2) regulates several key physiological and pathophysiological processes, and its dysregulation has been implicated in obesity, diabetes, and cancer. CaMKK2 is inhibited through phosphorylation in a process involving binding to the scaffolding 14–3–3 protein, which maintains CaMKK2 in the phosphorylation-mediated inhibited state. The previously reported structure of the N-terminal CaMKK2 14–3–3-binding motif bound to 14–3–3 suggested that the interaction between 14–3–3 and CaMKK2 could be stabilized by small-molecule compounds. Thus, we investigated the stabilization of interactions between CaMKK2 and 14–3–3γ by Fusicoccin A and other fusicoccanes—diterpene glycosides that bind at the interface between the 14–3–3 ligand binding groove and the 14–3–3 binding motif of the client protein. Our data reveal that two of five tested fusicoccanes considerably increase the binding of phosphopeptide representing the 14–3–3 binding motif of CaMKK2 to 14–3–3γ. Crystal structures of two ternary complexes suggest that the steric contacts between the C-terminal part of the CaMKK2 14–3–3 binding motif and the adjacent fusicoccane molecule are responsible for differences in stabilization potency between the study compounds. Moreover, our data also show that fusicoccanes enhance the binding affinity of phosphorylated full-length CaMKK2 to 14–3–3γ, which in turn slows down CaMKK2 dephosphorylation, thus keeping this protein in its phosphorylation-mediated inhibited state. Therefore, targeting the fusicoccin binding cavity of 14–3–3 by small-molecule compounds may offer an alternative strategy to suppress CaMKK2 activity by stabilizing its phosphorylation-mediated inhibited state.
Workplace	Institute of Physiology
Contact	Lucie Trajhanová, lucie.trajhanova@fgu.cas.cz, Tel.: 241 062 400
Year of Publishing	2021
Electronic address	https://doi.org/10.1021/acschembio.0c00821

Number of the records: 1