Prostaglandin H synthase kinetics in the two-phase aqueous-micellar system

Ponomareva, O.A.; Trushkin, N.A.; Filimonov, I.S.; Krivoshey, A.V.; Barkhatov, V.I.; Mitrofanov, S.I.; Vrzheshch, P.V.

Статья опубликована в высокорейтинговом журнале

Информация о цитировании статьи получена из Web of Science, Scopus
Статья опубликована в журнале из списка Web of Science и/или Scopus
Дата последнего поиска статьи во внешних источниках: 20 января 2019 г.

Авторы: Ponomareva Olga A., Trushkin Nikita A., Filimonov Ivan S., Krivoshey Alexandr V., Barkhatov Vladimir I., Mitrofanov Sergey I., Vrzheshch Petr V.
Журнал: Biochimica et Biophysica Acta - Biomembranes
Том: 1858
Номер: 9
Год издания: 2016
Издательство: Elsevier BV
Местоположение издательства: Netherlands
Первая страница: 2199
Последняя страница: 2207
DOI: 10.1016/j.bbamem.2016.06.013
Аннотация: Reaction mixture for PGHS (prostaglandin-H-synthase) is a two-phase system including micellar hydrophobic phase and hydrophilic aqueous phase. Reagents added to the mixture are distributed between phases, thus concentrations of reagents dissolved in phases can differ significantly from their overall contents. Using dynamic light scattering we found that the hydrophobic phase produced by tween-20 consists of micelles, which radius (4–5 nm) does not depend on either tween-20 overall content (0.1%–1%v/v) or arachidonic acid (AA) addition (10–1000 μM) or PGHS addition (1 μM). Tween-20 overall content changing from 0.1% to 2%v/v dramatically affected COX kinetic, but accounting AA distribution between phases allowed us to estimate “true” parameters, independent of the tween-20 overall content and the concentration of another substrate: KMOx equals 9.8 μM O2 in the aqueous phase or 0.0074 bar in the gaseous phase, KMAA equals 5400 μM AA in the phase of tween-20 micelles and 5400/P μM AA in the aqueous phase (P is the distribution ratio for the AA between the aqueous phase and the hydrophobic phase (P > > 1000)). This approach allowed to evaluate PS, the distribution ratio for the AA between the hydrophobic phase and the PGHS active center (PS ~ 310). This coefficient indicates the AA selectivity toward the cyclooxygenase active center.
Добавил в систему: Митрофанов Сергей Игоревич

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Prostaglandin H synthase kinetics in the two-phase aqueous-micellar systemстатья

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