Publication details

14-3-3ζ phosphomimicking mutants: an alternative to phosphorylation?

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Year of publication 2021
Type Article in Proceedings
Conference Študentská vedecká konferencia PriF UK 2021: zborník recenzovaných príspevkov
MU Faculty or unit

Central European Institute of Technology

Keywords 14-3-3; 14-3-3zeta; phosphomimicking mutant; monomerization; thermal stability
Description The 14-3-3 proteins are typically found in dimeric form crucial for their proper function in cells. Yet, a unique role of monomers has been debated in recent studies. Monomerization of 14-3-3? is regulated via phosphorylation of Ser58 located at dimer interface. However, it is difficult to obtain 14-3-3? specifically phosphorylated at Ser58 (pS58) in sufficient purity and amount. For these reasons three distinct phosphomimicking mutants, i. e. 14-3-3? S58E, S58D and S57D S58D were designed and studied in terms of the oligomeric nature and thermodynamical properties. Finally, they were compared to 14-3-3? wild type and monomeric mutant L12E M78K, used to approximate monomeric 14-3-3? pS58, to discuss whether they could be valid alternative for study of 14-3-3? pS58. Although phosphomimicking mutants S58E and S58D and 14-3-3? pS58 share negative charge at position 58, they were observed to exhibit distinct properties than monomeric 14-3-3?. Designed double mutant S57D S58D with additional negative charge in the Ser58 surrounding appeared as more appropriate alternative of pS58 protein.
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