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

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F67985823%3A_____%2F20%3A00535685" target="_blank" >RIV/67985823:_____/20:00535685 - isvavai.cz</a>

Nalezeny alternativní kódy

RIV/00216208:11310/20:10420687

Výsledek na webu

<a href="https://doi.org/10.1021/acschembio.0c00821" target="_blank" >https://doi.org/10.1021/acschembio.0c00821</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1021/acschembio.0c00821" target="_blank" >10.1021/acschembio.0c00821</a>

Jazyk výsledku

angličtina

Název v původním jazyce

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

Popis výsledku v původním jazyce

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.

Název v anglickém jazyce

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

Popis výsledku anglicky

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.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

10608 - Biochemistry and molecular biology

Projekt

<a href="/cs/project/GA19-00121S" target="_blank" >GA19-00121S: Molekulární podstata regulace proteinkinasové aktivity prostřednictvím proteinů 14-3-3</a><br>

Návaznosti

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Rok uplatnění

2020

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

ACS Chemical Biology

ISSN

1554-8929

e-ISSN

1554-8937

Svazek periodika

15

Číslo periodika v rámci svazku

11

Stát vydavatele periodika

US - Spojené státy americké

Počet stran výsledku

12

Strana od-do

3060-3071

Kód UT WoS článku

000592981100024

EID výsledku v databázi Scopus

2-s2.0-85096508476