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The Denaturant- and Mutation-Induced Disassembly of Pseudomonas aeruginosa Hexameric Hfq Y55W Mutantстатья
- Авторы: Marchenkov Victor, Lekontseva Natalia, Marchenko Natalia, Kashparov Ivan, Murina Victoriia, Nikulin Alexey, Filimonov Vladimir, Semisotnov Gennady
- Журнал: Molecules
- Том: 27
- Номер: 12
- Год издания: 2022
- Издательство: MDPI
- Местоположение издательства: Basel, Switzerland
- Первая страница: 3821
- DOI: 10.3390/molecules27123821
- Аннотация: Although oligomeric proteins are predominant in cells, their folding is poorly studied atpresent. This work is focused on the denaturant- and mutation-induced disassembly of the hexamericmutant Y55W of the Q host factor (Hfq) from mesophilic Pseudomonas aeruginosa (Pae). Usingintrinsic tryptophan fluorescence, dynamic light scattering (DLS), and high-performance liquidchromatography (HPLC), we show that the dissociation of Hfq Y55W occurs either under the effectof GuHCl or during the pre-denaturing transition, when the protein concentration is decreased, withboth events proceeding through the accumulation of stable intermediate states. With an extremely lowpH of 1.4, a low ionic strength, and decreasing protein concentration, the accumulated trimers anddimers turn into monomers. Also, we report on the structural features of monomeric Hfq resultingfrom a triple mutation (D9A/V43R/Y55W) within the inter-subunit surface of the protein. Thisglobular and rigidly packed monomer displays a high thermostability and an oligomer-like contentof the secondary structure, although its urea resistance is much lower.
- Добавил в систему: Семисотнов Геннадий Васильевич