Phosphorylated and Phosphomimicking Variants May Differ—A Case Study of 14-3-3 Protein

Result code in IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216224%3A14740%2F22%3A00119696" target="_blank" >RIV/00216224:14740/22:00119696 - isvavai.cz</a>

Result on the web

<a href="https://www.frontiersin.org/articles/10.3389/fchem.2022.835733/full" target="_blank" >https://www.frontiersin.org/articles/10.3389/fchem.2022.835733/full</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.3389/fchem.2022.835733" target="_blank" >10.3389/fchem.2022.835733</a>

Result language

angličtina

Original language name

Phosphorylated and Phosphomimicking Variants May Differ—A Case Study of 14-3-3 Protein

Original language description

Protein phosphorylation is a critical mechanism that biology uses to govern cellular processes. To study the impact of phosphorylation on protein properties, a fully and specifically phosphorylated sample is required although not always achievable. Commonly, this issue is overcome by installing phosphomimicking mutations at the desired site of phosphorylation. 14-3-3 proteins are regulatory protein hubs that interact with hundreds of phosphorylated proteins and modulate their structure and activity. 14-3-3 protein function relies on its dimeric nature, which is controlled by Ser58 phosphorylation. However, incomplete Ser58 phosphorylation has obstructed the detailed study of its effect so far. In the present study, we describe the full and specific phosphorylation of 14-3-3ζ protein at Ser58 and we compare its characteristics with phosphomimicking mutants that have been used in the past (S58E/D). Our results show that in case of the 14-3-3 proteins, phosphomimicking mutations are not a sufficient replacement for phosphorylation. At physiological concentrations of 14-3-3ζ protein, the dimer-monomer equilibrium of phosphorylated protein is much more shifted towards monomers than that of the phosphomimicking mutants. The oligomeric state also influences protein properties such as thermodynamic stability and hydrophobicity. Moreover, phosphorylation changes the localization of 14-3-3ζ in HeLa and U251 human cancer cells. In summary, our study highlights that phosphomimicking mutations may not faithfully represent the effects of phosphorylation on the protein structure and function and that their use should be justified by comparing to the genuinely phosphorylated counterpart.

Czech name

—

Czech description

—

Type

J_imp - Article in a specialist periodical, which is included in the Web of Science database

CEP classification

—

OECD FORD branch

10608 - Biochemistry and molecular biology

Project

Result was created during the realization of more than one project. More information in the Projects tab.

Continuities

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)<br>S - Specificky vyzkum na vysokych skolach

Publication year

2022

Confidentiality

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

Name of the periodical

Frontiers in Chemistry

ISSN

2296-2646

e-ISSN

2296-2646

Volume of the periodical

10

Issue of the periodical within the volume

March

Country of publishing house

CH - SWITZERLAND

Number of pages

17

Pages from-to

1-17

UT code for WoS article

000777334200001

EID of the result in the Scopus database

2-s2.0-85127127424