Russian Journal of Biological Physics and Chemisrty Russian Journal of Biological Physics and Chemisrty АКТУАЛЬНЫЕ ВОПРОСЫ БИОЛОГИЧЕСКОЙ ФИЗИКИ И ХИМИИ 2499-9962 54432 МЕДИЦИНСКАЯ БИОФИЗИКА И БИОФИЗИЧЕСКАЯ ХИМИЯ MEDICAL BIOPHYSICS AND BIOPHYSICAL CHEMISTRY МЕДИЦИНСКАЯ БИОФИЗИКА И БИОФИЗИЧЕСКАЯ ХИМИЯ EFFECTS OF TEMPERATURE ON CONFORMATIONAL DYNAMIC OF RED BLOOD CELLS AND INTACT HEMOGLOBIN ДИНАМИКА ИЗМЕНЕНИЯ КОНФОРМАЦИИ СВОБОДНОГО И ВНУТРИКЛЕТОЧНОГО ГЕМОГЛОБИНА ПРИ ТЕМПЕРАТУРНОМ ВОЗДЕЙСТВИИ Слатинская О. В. Slatinskaya O. V. slatolya@mail.ru Максимов Г. В. Maksimov G. V. Московский государственный университет им. М.В. Ломоносова Москва Россия Lomonosov Moscow State University Moscow Russian Federation Московский государственный университет им. М.В. Ломоносова Москва Россия Lomonosov Moscow State University Moscow Russian Federation 25 06 2019 25 06 2019 4 2 283 288 20 06 2019 20 06 2019 https://rusjbpc.ru/en/nauka/article/54432/view

В работе рассматривается изменение конформации гемоглобина в эритроцитах и выделенного гемоглобина в условиях нормоксии и гипоксии. Диффузия молекул кислорода и оксида азота через плазматическую мембрану эритроцита зависит от упорядоченности жирнокислотных хвостов фосфолипидов плазматической мембраны и состояния цитоскелета эритроцита, а способность гемоглобина связывать и сбрасывать эти молекулы зависит от его локализации в примембранной области и цитоплазме клетки. Исследование молекулярно-клеточных свойств эритроцитов, важных для осуществления их кислород-транспортной функции возможно с помощью метода спектроскопии комбинационного рассеяния (Раман-спектроскопия), при помощи которого возможно проводить регистрацию конформационных изменений гемоглобина эритроцитов in vitro и in vivo . В работе рассмотрены структурные изменения гема гемоглобина при взаимодействии с лигандами (О2 или СО2) при температурном воздействии. Использование лазера с длиной возбуждающего света 532 нм позволило изучить изменения в высокочастотной области спектра (2800-3100 см-1), характерной для изменения симметричных колебаний молекулярных связей -С-Н- метиленовых групп аминокислот глобина, что позволит установить взаимодействия между гемовой и глобиновой частью белка внутриклеточного и свободного гемоглобина. Соотношения полос спектра комбинационного рассеяния позволяют характеризовать динамику изменений конформации гема оксигемоглобна и дезоксигемоглобина при температурном воздействии в диапазоне от 25 до 36 °С. Показано, что изменения конформации различны для молекул гемоглобина в эритроцитах и свободного гемоглобина при температурном и гипоксическом воздействии, но не коррелируют с удельной растворимостью кислорода в растворе.

The conformational and structure changes of heme side-chains of human hemoglobin in red blood cells and intact hemoglobin upon ligand (O2 or CO2) dissociation have been studied be resonance Raman spectroscopy. Resonance Raman spectra of oxygenated and deoxygenated functional red blood cells and isolate hemoglobin recorded using 532 nm laser excitation are presented. The high sensitivity of Raman spectroscopy allows to registration conformation changes of heme (protoporphyrin) and globin part of protein in vivo and in vitro . We studied the effects of temperature on conformational dynamic changes of intracellular and intact hemoglobin upon hypoxia and normoxia states. We showed modification in the hemoglobin vibration bands in the CH2/CH3 stretching bands in the 2800-3100 cm-1 region. Spectral profile in the higher spectral region have been assigned to the symmetric / asymmetric CH, stretch, and symmetric / asymmetric CH3, stretch. This study provides new insights into the vibrational dynamics of heme and globin parts within functional red blood cells and solve to cooperative ligand binding mechanism of conformation changes of oxyhemoglobin and deoxyhemoglobin. Ratios from chosen Raman peaks were calculated and compared between the different subjects. We show that conformational and side-chains of hemoglobin in red blood cells and intact hemoglobin do not correlated with oxygen water solubility.

 гемоглобин эритроцит комбинационное рассеяние hemoglobin red blood cells Raman spectroscopy

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