Russian Journal of Biological Physics and Chemisrty Russian Journal of Biological Physics and Chemisrty АКТУАЛЬНЫЕ ВОПРОСЫ БИОЛОГИЧЕСКОЙ ФИЗИКИ И ХИМИИ 2499-9962 54641 МЕДИЦИНСКАЯ БИОФИЗИКА И БИОФИЗИЧЕСКАЯ ХИМИЯ MEDICAL BIOPHYSICS AND BIOPHYSICAL CHEMISTRY МЕДИЦИНСКАЯ БИОФИЗИКА И БИОФИЗИЧЕСКАЯ ХИМИЯ Proposed mechanism of regulation of the physiological effects of nitric oxide Предполагаемый механизм регуляции физиологических эффектов оксида азота Титов В. Ю. Titov V. Yu. vtitov43@yandex.ru Осипов А. Н. Osipov A. N. Кочиш И. И. Kochish I. I. kochish.i@mail.ru Российский национальный исследовательский медицинский университет имени Н.И. Пирогова Москва Россия Pirogov Russian National Research Medical University Moscow Russian Federation Московская государственная академия ветеринарной медицины и биотехнологии – МВА имени К.И. Скрябина Москва Россия Moscow State Academy of Veterinary Medicine and Biotechnology - MVA by K.I. Skryabin Moscow Russian Federation ФНЦ Всероссийский научно-исследовательский и технологический институт птицеводства РАН Сергиев Посад Россия Russian National Research and Technological Poultry Farming Institute, RAS Sergiev Posad Russian Federation Российский национальный исследовательский медицинский университет имени Н.И. Пирогова Москва Россия Pirogov Russian National Research Medical University Moscow Russian Federation Московская государственная академия ветеринарной медицины и биотехнологии - МВА имени К.И. Скрябина Россия Moscow state Academy of Veterinary Medicine and Biotechnology - MVA by K.I. Skryabin Russian Federation 25 06 2021 25 06 2021 6 2 289 295 20 06 2021 20 06 2021 https://rusjbpc.ru/en/nauka/article/54641/view

При помощи высокочувствительного и высокоспецифичного ферментного сенсора показано, что в норме большинство живых тканей содержит не более 50-100 нМ нитрита и нитрозоаминов, но единицы и десятки микромоль соединений - доноров NO: S-нитрозотиолов (RSNO), динитрозильных комплексов железа (ДНКЖ), высокомолекулярные нитросоединения, способные трансформироваться в ДНКЖ (RNO2). Следовательно, в норме живые ткани имеют механизмы предотвращения окисления NO кислородом до токсических продуктов. Доноры NO - стабильные соединения и, практически, не распадаются спонтанно с высвобождением NO. Основной пул доноров NO в большинстве тканей представлен ДНКЖ. NO может переходить с комплекса на мишень в момент деструкции комплекса под действием более эффективных хелаторов железа, чем лиганды, входящие в состав комплекса. Причем переход осуществляется с минимальным пребыванием NO в свободном состоянии. В случае, если комплекс подвергается воздействию эффективного хелатора железа, но мишень отсутствует, образуется железо-нитрозильный комплекс, содержащий этот хелатор. Мы предполагаем, что части апофермента некоторых ферментов -физиологических мишеней NO могут выступать в роли хелаторов - конкурентов. Таким образом, физиологический эффект соединений - доноров NO зависит не от их способности диссоциировать с высвобождением NO, но, прежде всего от наличия и состояния физиологической мишени. Не NO случайно находит мишень, а мишень, взаимодействует с донором - NO, вызывая его деструкцию и присоединяя NO. Также эффективность ДНКЖ как донора NO зависит от состава комплекса. Комплекс, содержащий лиганды с высоким сродством к железу, труднее разрушается хелатором железа, что необходимо для переноса NO к мишени. Это показано как на модельных системах, так и на живых организмах.

By means of highly sensitive and specific enzyme sensor it was shown, that most of living tissues contain less than 50-100 nM of nitrite and nitrosamines and up to tens of μM of NO donors - S-nitrosothiols (RSNO), dinitrosyl iron complexes (DNIC) and high molecular weight nitrocompounds, that can turn to DNIC (RNO2). This fact means that living tissues can prevent NO oxidation to produce toxic compounds. NO donors - are stable compounds and they are not spontaneously decomposed with free NO release. The main pool of NO donors in most tissues is represented by DNIC. NO can transfer from the complex to the target at the moment of the complex destruction under the action of more effective iron chelators than the ligands of the DNIC. The transition is carried out with a minimum stay of NO in the free state. If the complex is exposed to an effective iron chelator, but there is no target, an iron-nitrosyl complex containing this chelator is formed. We assume, that some parts of the apoenzymes of NO targets can play the role of competitive chelators. Therefore, the physiological effect of NO donors depend not on their ability to produce free NO, but on the presence and characteristics of the physiological target. Not free NO accidentally finds the target, but the target, interacts with the NO donor causing its destruction and attaching NO. The effectiveness of DNIC as NO donor also depends on the DNIC structure and the ligand type. DNIC containing ligands with a high affinity for iron is more difficult to destroy by an iron chelator, which is necessary for the transfer of NO to the target. These effects were demonstrated both in model systems and living organisms.

 Оксид азота динитрозильный комплекс железа (ДНКЖ) лиганды ДНКЖ nitric oxide dinitrosyl iron complexes (DNIC) DNIC ligands Исследование выполнено при финансовой поддержке РФФИ в рамках научного проекта 20-016-00204-а.

Vanin A., Borodulin R., Mikoyan V. Dinitrosyl iron complexes with natural thiol-containing ligands in aqueous solutions: Synthesis and some physico-chemical characteristics (A methodological review). Nitric Oxide., 2017, vol. 66, pp. 1-9. doi: 10.1016/j.niox.2017.02.005. Vanin A., Borodulin R., Mikoyan V. Dinitrosyl iron complexes with natural thiol-containing ligands in aqueous solutions: Synthesis and some physico-chemical characteristics (A methodological review). Nitric Oxide., 2017, vol. 66, pp. 1-9. doi: 10.1016/j.niox.2017.02.005. Vanin A. Dinitrosyl iron complexes with thiol-containing ligands as a "working form" of endogenous nitric oxide. Nitric Oxide, 2016, vol. 54, pp. 15-29. doi: 10.1016/j.niox.2016.01.006 Vanin A. Dinitrosyl iron complexes with thiol-containing ligands as a "working form" of endogenous nitric oxide. Nitric Oxide, 2016, vol. 54, pp. 15-29. doi: 10.1016/j.niox.2016.01.006 Hickok J.R., Sahni S., Shen H., Arvind A., Antoniou C., Fung L.W., Thomas D. Dinitrosyliron complexes are the most abundant nitric oxide-derived cellular adduct: biological parameters of assembly and disappearance. Free Radic. Biol. Med., 2011, vol. 51, no. 8, pp. 1558-1566. doi: 10.1016/j.freeradbiomed.2011.06.030 Hickok J.R., Sahni S., Shen H., Arvind A., Antoniou C., Fung L.W., Thomas D. Dinitrosyliron complexes are the most abundant nitric oxide-derived cellular adduct: biological parameters of assembly and disappearance. Free Radic. Biol. Med., 2011, vol. 51, no. 8, pp. 1558-1566. doi: 10.1016/j.freeradbiomed.2011.06.030 Tarpey M., Wink D., Grisham M., Methods for detection of reactive metabolites of oxygenand nitrogen: in vitro and in vivo considerations. American Journal of Physiology-Regulatory Integrative and Comparative Physiology, 2004, vol. 286, pp. R431-R444. doi:10.1152/ajpregu.00361.2003. Tarpey M., Wink D., Grisham M., Methods for detection of reactive metabolites of oxygenand nitrogen: in vitro and in vivo considerations. American Journal of Physiology-Regulatory Integrative and Comparative Physiology, 2004, vol. 286, pp. R431-R444. doi:10.1152/ajpregu.00361.2003. Titov V., Osipov A. Nitrite and Nitroso Compounds can serve as Specific Catalase Inhibitors. Redox Rep, 2017, vol. 22, no. 2, pp. 91-97. doi: 10.1080/13510002.2016.1168589. Titov V., Osipov A. Nitrite and Nitroso Compounds can serve as Specific Catalase Inhibitors. Redox Rep, 2017, vol. 22, no. 2, pp. 91-97. doi: 10.1080/13510002.2016.1168589. Titov V. The Enzymatic Technologies Open New Possibilities for Studying Nitric Oxide (NO) Metabolism in Living Systems. Current Enzyme Inhibition, 2011, vol. 7, no. 1, pp. 56-70. doi: 10.2174/157340811795713774. Titov V. The Enzymatic Technologies Open New Possibilities for Studying Nitric Oxide (NO) Metabolism in Living Systems. Current Enzyme Inhibition, 2011, vol. 7, no. 1, pp. 56-70. doi: 10.2174/157340811795713774. Severina I., Bussygina O., Pyatakova N., Malenkova I., Vanin A. Activation of soluble guanylate cyclase by NO donors--S-nitrosothiols, and dinitrosyl-iron complexes with thiol-containing ligands. Nitric Oxide, 2003, vol. 8, pp. 155-163. doi: 10.1016/s1089-8603(03)00002-8 Severina I., Bussygina O., Pyatakova N., Malenkova I., Vanin A. Activation of soluble guanylate cyclase by NO donors--S-nitrosothiols, and dinitrosyl-iron complexes with thiol-containing ligands. Nitric Oxide, 2003, vol. 8, pp. 155-163. doi: 10.1016/s1089-8603(03)00002-8 Titov V.Y., Kosenko O.V., Starkova E.S., Kondratov G.V., Borkhunova E.N., Petrov V.A., Osipov A.N. Enzymatic Sensor Detects Some Forms of Nitric Oxide Donors Undetectable by Other Methods in Living Tissues. Bull Exp Biol Med., 2016, vol. 162, no. 1, pp. 107-110. doi: 10.1007/s10517-016-3557-1. Titov V.Y., Kosenko O.V., Starkova E.S., Kondratov G.V., Borkhunova E.N., Petrov V.A., Osipov A.N. Enzymatic Sensor Detects Some Forms of Nitric Oxide Donors Undetectable by Other Methods in Living Tissues. Bull Exp Biol Med., 2016, vol. 162, no. 1, pp. 107-110. doi: 10.1007/s10517-016-3557-1. Титов В.Ю., Петренко Ю.М., Петров В.А., Владимиров Ю.А. Механизм окисления оксигемоглобина, индуцированного перекисью водорода. Бюлл. экспер. биол. мед., 1991, том 112, № 7, с. 46-49. @@Titov V., Petrenko Yu., Petrov V., Vladimirov Yu. Mechanism of oxyhemoglobin oxidation induced by hydrogen peroxide. Biull Eksp Biol Med., 1991, vol. 112, no. 7, pp. 46-49. (In Russ) Titov V.Yu., Petrenko Yu.M., Petrov V.A., Vladimirov Yu.A. Mehanizm okisleniya oksigemoglobina, inducirovannogo perekis'yu vodoroda. Byull. eksper. biol. med., 1991, tom 112, № 7, s. 46-49. @@Titov V., Petrenko Yu., Petrov V., Vladimirov Yu. Mechanism of oxyhemoglobin oxidation induced by hydrogen peroxide. Biull Eksp Biol Med., 1991, vol. 112, no. 7, pp. 46-49. (In Russ) Titov V., Dolgorukova A., Fisinin V., Borkhunova Ye., Kondratov G., Slesarenko N., Kochish I. The role of nitric oxide (NO) in the body growth rate of birds. World Poultry Science Journal, 2018, vol. 74, pp. 675-686. doi: org/10.1017/S0043933918000661 Titov V., Dolgorukova A., Fisinin V., Borkhunova Ye., Kondratov G., Slesarenko N., Kochish I. The role of nitric oxide (NO) in the body growth rate of birds. World Poultry Science Journal, 2018, vol. 74, pp. 675-686. doi: org/10.1017/S0043933918000661 Stalmer J., Singel D., Loscalzo, J. Biochemistry of nitric oxide and its redox-activated forms. Science, 1992, vol. 258, pp. 1898-1902. doi: 10.1126/science.1281928. Stalmer J., Singel D., Loscalzo, J. Biochemistry of nitric oxide and its redox-activated forms. Science, 1992, vol. 258, pp. 1898-1902. doi: 10.1126/science.1281928. Rossig L., Fichtlscherer B., Breitschopf K., Haendeler J., Zeiher A., Mulsch A., Dimmeler S. Nitric oxide inhibits caspase-3 by S-nitrosation in vivo. J. Biol. Chem., 1999, vol. 274, no. 11, p.6823-6826. doi: 10.1074/jbc.274.11.6823. Rossig L., Fichtlscherer B., Breitschopf K., Haendeler J., Zeiher A., Mulsch A., Dimmeler S. Nitric oxide inhibits caspase-3 by S-nitrosation in vivo. J. Biol. Chem., 1999, vol. 274, no. 11, p.6823-6826. doi: 10.1074/jbc.274.11.6823. Dimmeler S., Haendeler J., Nehls, M., Zeiher A. Suppression of apoptosis by nitric oxide via inhibition of interleukin-1beta-converting enzyme (ICE)-like and cysteine protease protein (CPP)-32-like proteases. J. Exp. Med., 1997, vol. 185, no. 4, pp. 601-607. doi: 10.1084/jem.185.4.601. Dimmeler S., Haendeler J., Nehls, M., Zeiher A. Suppression of apoptosis by nitric oxide via inhibition of interleukin-1beta-converting enzyme (ICE)-like and cysteine protease protein (CPP)-32-like proteases. J. Exp. Med., 1997, vol. 185, no. 4, pp. 601-607. doi: 10.1084/jem.185.4.601. Kim Y-M., Chung H-T., Simmons R., Billiar T. Cellular non-heme iron content is a determinant of nitric oxide-mediated apoptosis, necrosis, and caspase inhibition. J. Biol. Chem., 2000, vol. 275, no. 15, pp. 10954-10961. doi: 10.1074/jbc.275.15.10954. Kim Y-M., Chung H-T., Simmons R., Billiar T. Cellular non-heme iron content is a determinant of nitric oxide-mediated apoptosis, necrosis, and caspase inhibition. J. Biol. Chem., 2000, vol. 275, no. 15, pp. 10954-10961. doi: 10.1074/jbc.275.15.10954. Schopfer F., Baker P., Giles G., Chumley Ph., Batthyany C., Crawford J., Patel R., Hogg N., Branchaud B., Lancaster J., Freeman B. Fatty acid transduction of nitric oxide signaling. Nitrolinoleic acid is a hydrophobically stabilized nitric oxide donor. J. Biol. Chem., 2005, vol. 280, no. 19, pp. 19289-19297. doi: 10.1074/jbc.M414689200 Schopfer F., Baker P., Giles G., Chumley Ph., Batthyany C., Crawford J., Patel R., Hogg N., Branchaud B., Lancaster J., Freeman B. Fatty acid transduction of nitric oxide signaling. Nitrolinoleic acid is a hydrophobically stabilized nitric oxide donor. J. Biol. Chem., 2005, vol. 280, no. 19, pp. 19289-19297. doi: 10.1074/jbc.M414689200 Lima E., Bonini M., Augusto O., Barbeiro H., Souza H., Abdalla D. Nitrated lipids decompose to nitric oxide and lipid radicals and cause vasorelaxation. Free Radic. Biol. & Med., 2005, vol. 39, no. 4, pp. 532-539. doi: 10.1016/j.freeradbiomed Lima E., Bonini M., Augusto O., Barbeiro H., Souza H., Abdalla D. Nitrated lipids decompose to nitric oxide and lipid radicals and cause vasorelaxation. Free Radic. Biol. & Med., 2005, vol. 39, no. 4, pp. 532-539. doi: 10.1016/j.freeradbiomed Forman H., Zhang H., Rinna A. Glutathione: Overview of its protective roles, measurement, and biosynthesis. Mol. Aspects Med., 2009, vol. 30, no. 1-2, pp. 1-12. doi: 10.1016/j.mam.2008.08.006 Forman H., Zhang H., Rinna A. Glutathione: Overview of its protective roles, measurement, and biosynthesis. Mol. Aspects Med., 2009, vol. 30, no. 1-2, pp. 1-12. doi: 10.1016/j.mam.2008.08.006