danДоклады Национальной академии наук БеларусиDoklady of the National Academy of Sciences of Belarus1561-83232524-2431The Republican Unitary Enterprise Publishing House "Belaruskaya Navuka"10.29235/1561-8323-2018-62-1-51-65dan-489Research ArticleХИМИЯCHEMISTRYЛИГАНД-СВЯЗЫВАЮЩИЕ И КАТАЛИТИЧЕСКИЕ СВОЙСТВА РЕКОМБИНАНТНОЙ ТРОМБОКСАНСИНТАЗЫ ЧЕЛОВЕКАLIGAND-BINDING AND CATALYTIC PROPERTIES OF RECOMBINANT HUMAN THROMBOXANE SYNTHASEСвиридА. В.SviridAndrey V.

науч. сотрудник

ул. Купревича, 5/2, 220141, Минск

Researcher

5/2, Kuprevich Str., 220014, Minsk

svirid.andrei@gmail.comШапироМ. А.ShapiraMichail A.

мл. науч. сотрудник

ул. Купревича, 5/2, 220141, Минск

Junior Researcher

5/2, Kuprevich Str., 220014, Minsk

mshapira2016@gmail.comШагойкоП. Г.ShahoikaPavel G.

химик

аг. Лесной, 31, Минская об- ласть

Chemist

31, Lesnoy, Minsk Region

pavel11sg@gmail.comПоходняЮ. Г.PakhadniaYury G.

канд. биол. наук, начальник отдела

аг. Лесной, 31, Минская область

Ph. D. (Biology), Head of the Department

31, Lesnoy, Minsk Region

pohodnia@yahoo.comГилепА. А.GilepAndrei A.

канд. хим. наук, вед. науч. сотрудник

ул. Купревича, 5/2, 220141, Минск

Ph. D. (Chemistry), Leading researcher

5/2, Kuprevich Str., 220014, Minsk

agilep@iboch.bas-net.byУсановС. А.UsanovSergei A.

член-корреспондент, д-р хим. наук, профессор

ул. Купревича, 5/2, 220141, Минск

Corresponding Member, D. Sc. (Chemistry), Professor

5/2, Kuprevich Str., 220014, Minsk

usanov@iboch.bas-net.byИнститут биоорганической химии Национальной академии наук БеларусиInstitute of Bioorganic Chemistry of the National Academy of Sciences of BelarusНациональная антидопинговая лабораторияNational Antidoping Laboratory2018040320186215165Copyright © Свирид А.В., Шапиро М.А., Шагойко П.Г., Походня Ю.Г., Гилеп А.А., Усанов С.А., 20182018Свирид А.В., Шапиро М.А., Шагойко П.Г., Походня Ю.Г., Гилеп А.А., Усанов С.А.Svirid A.V., Shapira M.A., Shahoika P.G., Pakhadnia Y.G., Gilep A.A., Usanov S.A.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://doklady.belnauka.by/jour/article/view/489

С целью выявления модуляторов активности тромбоксансинтазы человека осуществлен анализ взаимодействия рекомбинантного гемопротеида с различными низкомолекулярными лигандами. Показано, что тромбоксансинтаза взаимодействует с рядом жирных кислот и их производных (потенциальные субстраты или конкурентные ингибиторы), а также является мишенью для ингибирования производными имидазола и триазола, применяющимися в медицинской практике и сельском хозяйстве, что позволило установить механизм, по которому могут реализовывать свое токсическое действие соединения, нарушающие работу эндокринной системы человека (endocrine-disrupting chemicals, EDC). Впервые показано восстановление атома железа гема тромбоксансинтазы NАDPH-цитохром P450 редуктазой, что оказывает частичное ингибирующее действие на фермент и подавляет образование побочных продуктов реакции – 12-гидроксигептадекатриеноевой кислоты (12-HHT) и малонового диальдегида (МДА). Предполагается, что данный механизм может участвовать в регуляции активности фермента in vivo. 

To study the spectrum of modulators of the human thromboxane synthase activity, the interaction of recombinant protein with various low-molecular weight ligands was analyzed. It was shown that thromboxane synthase interacts with a number of fatty acids and their derivatives (potential substrates or concurrent inhibitors), being a target for nonselective inhibition by imidazole and triazole derivatives used in medical practice and agriculture. Thus, another mechanism of action of endocrine-disrupting chemicals (EDC) was established. For the first time, the reduction of heme iron of thromboxane synthase by cytochrome P450 reductase was shown. This interaction accompanied by a partial inhibitory effect on the enzyme suppresses the formation of reaction by-products 12-hydroxyheptadecatenoic acid (12-HHT) and malonic dialdehyde (MDA). It is likely that this mechanism can participate in the regulation of the enzyme activity in vivo. 

тромбоксансинтаза человека (TXAS)разностное спектрофотометрическое титрованиеNАDPH-цитохром P450 редуктаза (CPR)азолыжирные кислотыendocrine-disrupting chemicals (EDC)human thromboxane synthase (TXAS)differential spectrophotometric titrationcytochrome P450 reductase (CPR)azolesfatty acidsendocrine-disrupting chemicals (EDC)Работа выполнялась при поддержке БРРФИ (грант Х16Р-062). Коллектив авто- ров выражает благодарность старшему научному сотруднику лаборатории белковой инженерии Я. В. Диченко за помощь в постановке in silico экспериментов.This work was supported by the Belarussian Foundation for Basic Research (grant no. Х16Р-062). Authors acknowledge Ya. V. Dichenko for assistance in carrying out in silico experiments.ReferencesUllrich, V. Prostacyclin and thromboxane synthase as P-450 enzymes / V. Ullrich, H. Graf // Trends Pharmacol. Sci. – 1984. – Vol. 5 – P. 352–355. doi.org/10.1016/0165-6147(84)90467-xUllrich V., Graf H. Prostacyclin and thromboxane synthase as P-450 enzymes. 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The authors declare that there are no conflicts of interest present.