vestibИзвестия Национальной академии наук Беларуси. Серия биологических наукProceedings of the National Academy of Sciences of Belarus, Biological Series1029-89402524-230XThe Republican Unitary Enterprise Publishing House "Belaruskaya Navuka"10.29235/1029-8940-2023-68-3-183-196vestib-882Research ArticleСтатьиРоль дисульфидных связей в формировании пространственной структуры эпидермального фактора роста человекаThe role of disulfide bonds in the formation of the spatial structure of the human epidermal growth factorАкуневичА. А.AkunevichA. A.

Акуневич Анастасия Александровна – аспирант.

Пр. Дзержинского, 83, 220083, Минск

Anastasia A. Akunevich – Postgraduate student.

83, Dzerzhinski Ave., 220083, Minsk

akunevichna@gmail.comХрусталёвВ. В.KhrustalevV. V.

Хрусталёв Владислав Викторович – доктор биологических наук, доцент, заведующий кафедрой.

Пр. Дзержинского, 83, 220083, Минск

Vladislav V. Khrustalev – D. Sc. (Biol.), Associate Professor, Head of the Department.

83, Dzerzhinski Ave., 220083, Minsk

vvkhrustalev@mail.ruХрусталёваТ. А.KhrustalevaT. A.

Хрусталёва Татьяна Александровна – кандидат биологических наук, учeный секретарь.

Ул. Академическая, 28, 220072, г. Минск

Tatyana A. Khrustaleva – Ph. D. (Biol.), Scientific Secretary.

28, Akademicheskaya Str., 220072, Minsk

tanissia.lir@gmail.comКордюковаЛ. В.KordyukovaL. V.

Кордюкова Лариса Валентиновна – доктор биологических наук, вед. науч. сотрудник.

Ленинские горы, 1-40, 119991, Москва

Larisa V. Kordyukova – D. Sc. (Biol.), Leading Researcher.

1-40, Leninskiye gory, 119991, Moscow

kord@belozersky.msu.ruАрутюнянА. М.ArutyunyanA. M.

Арутюнян Александр Мигранович – кандидат физико-математических наук, доцент, заведующий лабораторией.

Ленинские горы, 1-40, 119991, Москва

Alexander M. Arutyunyan – Ph. D. (Phys.-Math.), Associate Professor, Head of the Laboratory.

1-40, Leninskiye gory, 119991, Moscow

alarut@belozersky.msu.ruБелорусский государственный медицинский университетBelarusian State Medical UniversityИнститут физиологии НАН БеларусиInstitute of Physiology of the National Academy of Sciences of BelarusНаучно-исследовательский институт физико-химической биологии имени А.Н. Белозерского Московского государственного университета имени М.В. ЛомоносоваA.N. Belozerski Institute of Physico-Chemical Biology, Lomonosov Moscow State University202329072023683183196Copyright © Акуневич А.А., Хрусталёв В.В., Хрусталёва Т.А., Кордюкова Л.В., Арутюнян А.М., 20232023Акуневич А.А., Хрусталёв В.В., Хрусталёва Т.А., Кордюкова Л.В., Арутюнян А.М.Akunevich A.A., Khrustalev V.V., Khrustaleva T.A., Kordyukova L.V., Arutyunyan A.M.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://vestibio.belnauka.by/jour/article/view/882

Эпидермальный фактор роста (EGF) – пептид семейства EGF-подобных факторов роста с общим консервативным EGF-подобным доменом, формируемым тремя внутримолекулярными дисульфидными связями. В данной работе описаны изменения в пространственной структуре EGF и его мутантной форме с заменой D46G в C-концевом фрагменте, наблюдаемые при восстановлении дисульфидных связей соответствующих синтетических пептидов в 0,01 М фосфатном буфере (рН = 7,4). Для анализа структуры использовались спектроскопия кругового дихроизма, спектрофлуориметрия, нативный электрофорез в полиакриламидном геле, центрифужная ультрафильтрация. Показано, что восстановление дисульфидных связей изменяет геометрию EGF-подобного домена в сторону увеличения содержания бета-структуры при сохранении димерной формы пептидов. Согласно результатам молекулярного моделирования, при этом может произойти удлинение главной бета-шпильки EGF-подобного домена, межмолекулярной бета-структуры или образование новой бета-структуры между N- и C-концевым фрагментами молекулы, что изменит поверхность межмолекулярного контакта в составе димера. Доказано, что восстановление дисульфидных связей препятствует диссоциации димеров EGF до мономеров. В физиологических условиях это может привести к неспособности EGF формировать сайты связывания с рецептором EGFR и вызывать его активацию.

The epidermal growth factor (EGF) is a peptide of the EGF-like growth factor family with a common conserved EGF-like domain formed by three intramolecular disulfide bonds. This article describes changes in the spatial structure of EGF and its mutant form with the D46G substitution in its C-terminal fragment observed upon disulfide bonds reduction in the corresponding synthetic peptides in a 0.01 M phosphate buffer (pH = 7.4). The structure was analyzed using circular dichroism spectroscopy, spectrofluorimetry, native polyacrylamide gel electrophoresis, and centrifugal ultrafiltration. It was shown that disulfide bonds reduction changes the geometry of the EGF-like domain towards an increase in the content of the beta-structure, while these peptides remain in dimeric form. According to the molecular modeling results, this can lead to the elongation of the main beta-hairpin of the EGF-like domain, to the elongation of the intermolecular beta-structure, or to the formation of a new beta-structure between the N- and C-terminal fragments of each molecule, which will change the intermolecular interface in dimeric form. Disulfide bonds reduction prevents EGF dimer dissociation to monomers. Under physiological conditions, this can lead to the inability of EGF to form binding sites for EGFR (epidermal growth factor receptor) and to cause its activation.

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The authors declare that there are no conflicts of interest present.