Free energy calculations on the stability of the 14-3-3 zeta protein

Varování

Publikace nespadá pod Ekonomicko-správní fakultu, ale pod Středoevropský technologický institut. Oficiální stránka publikace je na webu muni.cz.
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JANDOVA Z. TROŠANOVÁ Zuzana WEISOVÁ Veronika OOSTENBRINK C. HRITZ Jozef

Rok publikování 2018
Druh Článek v odborném periodiku
Časopis / Zdroj Biochimica et Biophysica Acta - Proteins and Proteomics
Fakulta / Pracoviště MU

Středoevropský technologický institut

Citace
www https://www.sciencedirect.com/science/article/pii/S1570963917302807?via%3Dihub
Doi http://dx.doi.org/10.1016/j.bbapap.2017.11.012
Klíčová slova 14-3-3 protein; Protein stability; Molecular dynamics simulation; Differential scanning calorimetry; Free energy calculation; Thermodynamic integration
Popis Mutations of cysteine are often introduced to e.g. avoid formation of non-physiological inter-molecular disulfide bridges in in-vitro experiments, or to maintain specificity in labeling experiments. Alanine or serine is typically preferred, which usually do not alter the overall protein stability, when the original cysteine was surface exposed. However, selecting the optimal mutation for cysteines in the hydrophobic core of the protein is more challenging. In this work, the stability of selected Cys mutants of 14-3-3 zeta was predicted by free-energy calculations and the obtained data were compared with experimentally determined stabilities. Both the computational predictions as well as the experimental validation point at a significant destabilization of mutants C94A and C94S. This destabilization could be attributed to the formation of hydrophobic cavities and a polar solvation of a hydrophilic side chain. A L12E, M78K double mutant was further studied in terms of its reduced dimerization propensity. In contrast to naive expectations, this double mutant did not lead to the formation of strong salt bridges, which was rationalized in terms of a preferred solvation of the ionic species. Again, experiments agreed with the calculations by confirming the monomerization of the double mutants. Overall, the simulation data is in good agreement with experiments and offers additional insight into the stability and dimerization of this important family of regulatory proteins.
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