Fluorescence lifetime components reveal kinetic intermediate states upon equilibrium denaturation of carbonic anhydrase II

Описание

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.

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Издание

Журнал: METHODS AND APPLICATIONS IN FLUORESCENCE

Выпуск журнала: Vol. 6, Is. 1

ISSN журнала: 20506120

Место издания: BRISTOL

Издатель: IOP PUBLISHING LTD

Авторы

  • Nemtseva Elena V. (Siberian Fed Univ, Krasnoyarsk 660041, Russia; Krasnoyarsk Sci Ctr SB RAS, Fed Res Ctr, Inst Biophys SB RAS, Krasnoyarsk 660036, Russia)
  • Lashchuk Olesya O. (Siberian Fed Univ, Krasnoyarsk 660041, Russia)
  • Gerasimova Marina A. (Siberian Fed Univ, Krasnoyarsk 660041, Russia)
  • Melnik Tatiana N. (Russian Acad Sci, Inst Prot Res, Pushchino 142290, Moscow Region, Russia)
  • Nagibina Galina S. (Russian Acad Sci, Inst Prot Res, Pushchino 142290, Moscow Region, Russia)
  • Melnik Bogdan S. (Russian Acad Sci, Inst Prot Res, Pushchino 142290, Moscow Region, Russia)

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