Russian Journal of Biological Physics and Chemisrty Russian Journal of Biological Physics and Chemisrty АКТУАЛЬНЫЕ ВОПРОСЫ БИОЛОГИЧЕСКОЙ ФИЗИКИ И ХИМИИ 2499-9962 54342 МЕДИЦИНСКАЯ БИОФИЗИКА И БИОФИЗИЧЕСКАЯ ХИМИЯ MEDICAL BIOPHYSICS AND BIOPHYSICAL CHEMISTRY МЕДИЦИНСКАЯ БИОФИЗИКА И БИОФИЗИЧЕСКАЯ ХИМИЯ STRUCTURAL FEATURES AND STABILITY OF CARBONIC ANHYDRASE INTERMEDIATE STATES СТРУКТУРНЫЕ ОСОБЕННОСТИ И СТАБИЛЬНОСТЬ ПРОМЕЖУТОЧНЫХ СОСТОЯНИЙ КАРБОКСИАНГИДРАЗЫ Катина Н С Katina N S delfinium27@rambler.ru Балобанов В А Balobanov V A Ильина Н Б Ilyina N B Маркелова Н Ю Markelova N Y Кашпаров И А Kashparov I A Марченков В В Marchenkov V V Институт белка РАН ru Institute of Protein Research RAS ru Институт белка РАН ru Institute of Protein Research RAS ru Институт белка РАН ru Institute of Protein Research RAS ru Институт биофизики клетки РАН ru Institute of Cell Biophysics ru Институт белка РАН ru Institute of Protein Research RAS ru Институт белка РАН ru Institute of Protein Research RAS ru 25 09 2018 25 09 2018 3 3 642 650 20 09 2018 20 09 2018 https://rusjbpc.ru/en/nauka/article/54342/view

Карбоксиангидраза представляет собой однодоменный глобулярный белок, в нативном состоянии которого преобладает β-структура. В данной работе проведено исследование равновесного разворачивания апоформы карбоксиангидразы понижением рН. Показано, что денатурация белка кислым рН представляет собой переход между тремя состояниями N → I1→ I2. Добавление мочевины к I2 приводит к его разворачиванию до развернутого состояния I2 → U. С помощью аппроксимации переходов равновесного разворачивания белка понижением рН и мочевиной удалось провести оценку стабильностей нативного и двух промежуточных состояний карбоксиангидразы. Описанный подход может быть использован в дальнейшем для исследования влияния аминокислотных замен на сворачивание данного белка. Также в работе были исследованы структурные особенности конформационных состояний карбоксиангидразы, полученные результаты позволили сделать вывод, что состояние I1 находится в составе ассоциатов и содержит ненативные α-спиральные участки.

Carbonic anhydrase is a globular single-domain protein; its native state is dominated by β-structure. In this work the study of pH-induced equilibrium unfolding of carbonic anhydrase apoform has been carried out. It was shown that the protein denaturation by low pH represents the tree-state transition N → I1 → I2. Addition of urea to I2 leads to its unfolding to the unfolded state I2 → U. Using approximation of pH-induced and urea-induced equilibrium unfolding transition curves the estimation of the stability of carbonic anhydrase native and both intermediate states was carried out. Described approach can be used further for investigation of the influence of amino acid substitutions on the folding process of this protein. Moreover in this work the structural features of carbonic anhydrase conformational states were investigated, obtained results concluded that I1 forms associates and contains nonnative α-helical regions.

 карбоксиангидраза сворачивание белка промежуточное состояние равновесное разворачивание круговой дихроизм carbonic anhydrase protein folding intermediate state equilibrium unfolding circular dichroism

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