Тип публикации: статья из журнала
Год издания: 2018
Идентификатор DOI: 10.1088/2050-6120/aa994a
Ключевые слова: time-resolved fluorescence spectroscopy, carbonic anhydrase II, protein intermediate states, comparison of kinetic and equilibrium experiments, protein fluorescence lifetime
Аннотация: In most cases, intermediate states of multistage folding proteins are not 'visible' under equilibrium conditions but are revealed in kinetic experiments. Time-resolved fluorescence spectroscopy was used in equilibrium denaturation studies. The technique allows for detecting changes in the conformation and environment of tryptophan Показать полностьюresidues in different structural elements of carbonic anhydrase II which in its turn has made it possible to study the intermediate states of carbonic anhydrase II under equilibrium conditions. The results of equilibrium and kinetic experiments using wild-type bovine carbonic anhydrase II and its mutant form with the substitution of leucine for alanine at position 139 (L139A) were compared. The obtained lifetime components of intrinsic tryptophan fluorescence allowed for revealing that, the same as in kinetic experiments, under equilibrium conditions the unfolding of carbonic anhydrase II ensues through formation of intermediate states.
Журнал: METHODS AND APPLICATIONS IN FLUORESCENCE
Выпуск журнала: Vol. 6, Is. 1
Номера страниц: 15006
ISSN журнала: 20506120
Место издания: BRISTOL
Издатель: IOP PUBLISHING LTD