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Comparative stability of dihydrofolate reductase mutants in vitro and in vivoстатья
- Авторы: Leontiev Vladimir V., Uversky Vladimir N., Gudkov Anatoly T.
- Журнал: Protein engineering
- Том: 6
- Номер: 1
- Год издания: 1993
- Издательство: Oxford University Press
- Местоположение издательства: United Kingdom
- Первая страница: 81
- Последняя страница: 84
- DOI: 10.1093/protein/6.1.81
- Аннотация: Dihydrofolate reductase mutants with amino acid replacements in the active center (Thr35-->Asp mutant, Arg57-->His mutant and the mutant with triple replacement Thr35-->Asp, Asn37-->Ser, Arg57-->His) were obtained by site-directed mutagenesis. The stabilization effect of trimethoprim and NADP.H on the protein tertiary structure in vitro has been investigated. In the case of mutants with a 'weak' tertiary structure (Thr35-->Asp35 and the triple mutant) the separate addition of ligands does not affect their stability. The simultaneous addition of these ligands to Thr35-->Asp35 and the triple mutant leads to the large increase in their stability. A distinct correlation was found between the in vitro studied stability of the mutant proteins to the urea- or heat-induced denaturation and the level of proteolytic degradation of these mutants previously observed in vivo.
- Добавил в систему: Уверский Владимир Николаевич