Publication details

Phosphomimicking mutants: why to be careful



Year of publication 2020
Type Conference abstract
MU Faculty or unit

Central European Institute of Technology

Description Stability of protein is the ability to maintain its structure and thus function under stress. This stress can be caused by alternations of surroundings such as temperature or pH change, addition of denaturant, presence of salts, etc. [1]. Many methods to assess protein stability were developed, including nanoDSF, which measures changes in the intrinsic fluorescence of tryptophan during protein unfolding, to determine the melting temperature [2]. Phosphomimicking mutants are a tool commonly used to study the properties of phosphorylated proteins [3]. Here we compared thermodynamic stability of such mutant and phosphorylated form of 14-3-3? protein. This particular protein plays a role in regulation of many cellular functions [4]. The phosphorylation of S58 located at the dimer interface is one of the factors affecting interaction of 14-3-3? with its partners [5]. The results have shown, that phosphomimicking mutant S58E cannot be considered a sufficient replacement for phosphorylated 14-3-3? at S58. Not only does their stability differ in various conditions, with overall better stability of S58E, but their intrinsic fluorescence behaves differently as well.
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