Quantification of superoxide radical production in thylakoid membrane using cyclic hydroxylamines

Quantification of superoxide radical production in thylakoid membrane using cyclic hydroxylamines Научная публикация

Журнал

Free Radical Biology and Medicine
ISSN: 0891-5849

Вых. Данные

Год: 2015, Том: 89, Страницы: 1014-1023 Страниц : 10 DOI: 10.1016/j.freeradbiomed.2015.08.016

Ключевые слова

Reactive Oxygen Species; superoxide anion radical; photosynthetic electron transfer chain; cyclic hydroxylamines; electron spin resonance

Авторы

Kozuleva Marina ¹ , Klenina Irina ¹ , Mysin Ivan ^1,2 , Kirilyuk Igor ^3,4 , Opanasenko Vera ¹ , Proskuryakov Ivan ¹ , Ivanov Boris ¹

Организации

1	(Данные Web of science) Russian Acad Sci, Inst Basic Biol Problems, Pushchino 142290, Russia
2	(Данные Web of science) Russian Acad Sci, Inst Theoret & Expt Biophys, Pushchino 142290, Russia
3	(Данные Web of science) Russian Acad Sci, Siberian Branch, Novosibirsk Organ Chem Inst, Novosibirsk 630090, Russia
4	(Данные Web of science) Novosibirsk State Univ, Novosibirsk 630090, Russia

Applicability of two lipophilic cyclic hydroxylamines (CHAs), CM-H and TMT-H, and two hydrophilic CHAs, CAT1-H and DCP-H, for detection of superoxide anion radical (O-2(center dot-)) produced by the thylakoid photosynthetic electron transfer chain (PETC) of higher plants under illumination has been studied. ESR spectrometry was applied for detection of the nitroxide radical originating due to CHAs oxidation by O-2(center dot) CHAs and corresponding nitroxide radicals were shown to be involved in side reactions with PETC which could cause miscalculation of O-2(center dot-) production rate. Lipophilic CM-H was oxidized by PETC components, reducing the oxidized donor of Photosystem I, P-700(+) Not while at the same concentration another lipophilic CHA, TMT-H, did not reduce P-700(+) The nitroxide radical was able to accept electrons from components of the photosynthetic chain. Electrostatic interaction of stable cation CAT1-H with the membrane surface was suggested. Water-soluble superoxide dismutase (SOD) was added in order to suppress the reaction of CHA with O-2(center dot-) outside the membrane. SOD almost completely inhibited light-induced accumulation of DCP center dot, nitroxide radical derivative of hydrophilic DCP-H, in contrast to TMT center dot accumulation. Based on the results showing that change in the thylakoid lumen pH and volume had minor effect on TMT* accumulation, the reaction of TMT-H with O-2(center dot-) in the lumen was excluded. Addition of TMT-H to thylakoid suspension in the presence of SOD resulted in the increase in light-induced O-2 uptake rate, that argued in favor of TMT-H ability to detect O-2(center dot-) produced within the membrane core. Thus, hydrophilic DCP-H and lipophilic TMT-H were shown to be usable for detection of O-2(center dot-) produced outside and within thylakoid membranes. (C) 2015 Elsevier Inc. All rights reserved.

Kozuleva M. , Klenina I. , Mysin I. , Kirilyuk I. , Opanasenko V. , Proskuryakov I. , Ivanov B.
Quantification of superoxide radical production in thylakoid membrane using cyclic hydroxylamines
Free Radical Biology and Medicine. 2015. V.89. P.1014-1023. DOI: 10.1016/j.freeradbiomed.2015.08.016 WOS Scopus РИНЦ

Опубликована в печати:

1 дек. 2015 г.

Web of science	WOS:000366355800095
Scopus	2-s2.0-84946144365
РИНЦ	24968774
OpenAlex	W1872846668