danДоклады Национальной академии наук БеларусиDoklady of the National Academy of Sciences of Belarus1561-83232524-2431The Republican Unitary Enterprise Publishing House "Belaruskaya Navuka"10.29235/1561-8323-2021-65-5-568-575dan-1006Research ArticleХИМИЯCHEMISTRYДизайн структуры химерного белка ДНК-экзотрансферазы быка и SSB-белка E. coliDesign structure of fusion protein of bovine DNA exotransferase and E. coli SSB proteinСаченкоА. Б.SachankaA. B.

Саченко Антон Борисович – мл. науч. сотрудник, аспирант

ул. Купревича, 5/2, 220141, Минск

Sachanka Antos B. – Junior researcher, Postgraduate student

5/2, Kuprevich Str., 220141, Minsk

antosuk@yandex.ruДиченкоЯ. В.DzichenkaYa. U.

Диченко Ярослав Владимирович – канд. хим. наук, ст. науч. сотрудник

ул. Купревича, 5/2, 220141, Минск

Dzichenka Yaraslau U. – Ph. D. (Chemistry), Senior researcher

5/2, Kuprevich Str., 220141, Minsk

dichenko@iboch.byЯнцевичА. В.YantsevichA. V.

Янцевич Алексей Викторович – канд. хим. наук, заведующий лабораторией

ул. Купревича, 5/2, 220141, Минск

Yantsevich Aliaksei V. – Ph. D. (Chemistry), Head of the Laboratory

5/2, Kuprevich Str., 220141, Minsk

al.yantsevich@gmail.comУсановС. А.UsanovS. A.

Усанов Сергей Александрович – член-корреспондент, д-р хим. наук, профессор

ул. Купревича, 5/2, 220141, Минск

Usanov Sergei A. – Corresponding Member, D. Sc. (Chemistry), Professor

5/2, Kuprevich Str., 220141, Minsk

usanov@iboch.byИнститут биоорганической химии Национальной академии наук БеларусиInstitute of Bioorganic Chemistry of the National Academy of Sciences of Belarus202107112021655568575Copyright © Саченко А.Б., Диченко Я.В., Янцевич А.В., Усанов С.А., 20212021Саченко А.Б., Диченко Я.В., Янцевич А.В., Усанов С.А.Sachanka A.B., Dzichenka Y.U., Yantsevich A.V., 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/1006

С целью изучения влияния дополнительного ДНК-связывающего домена SSB-белка E. coli, присоединенного к транкированной и нативной ДНК-экзотрансферазе быка, на ДНК-аффинность и стабильность фермента, проведен анализ траекторий молекулярной динамики и пространственных структур гомологичных моделей химерного белка с различными линкерами. Установлено, что более предпочтительным для присоединения SSB-белка является C-концевая последовательность фермента, при этом прогнозируемая стабильность транкированного химерного фермента выше, чем у нативного. Согласно данным молекулярной динамики, введение линкеров между двумя белками для нативной (GGGGSGGGSGGGGS, GGGSGGGS и TCT) и транкированной (GGSGGGSGG, GGGGGG, GTGSGT и 5xGGGGS) формы фермента не только способствует повышению его стабильности, но и увеличивает взаимную подвижность ДНК-аффинных доменов.The analysis of the trajectories of molecular dynamics simulation and spatial structures of homologous models of fusion protein with various linkers was performed to understand the effect of the additional DNA-binding domain of the E. coli SSB protein attached to the truncated and native bovine DNA exotransferase on its stability and activity. It is found that the C-terminus of the enzyme is the preferred end for attachment of the E. coli protein, while the stability of the truncated fusion enzyme is higher than the native one. According to molecular dynamics data, introducing linkers between two proteins for the native (GGGGSGGGSGGGGS, GGGSGGGS, and TCT) and truncated (GGSGGGSGG, GGGGGG, GTGSGT, and 5xGGGGS) forms of the enzyme not only improves its stability, but also increases the mutual mobility of DNA-affinity domains.

The analysis of the trajectories of molecular dynamics simulation and spatial structures of homologous models of fusion protein with various linkers was performed to understand the effect of the additional DNA-binding domain of the E. coli SSB protein attached to the truncated and native bovine DNA exotransferase on its stability and activity. It is found that the C-terminus of the enzyme is the preferred end for attachment of the E. coli protein, while the stability of the truncated fusion enzyme is higher than the native one. According to molecular dynamics data, introducing linkers between two proteins for the native (GGGGSGGGSGGGGS, GGGSGGGS, and TCT) and truncated (GGSGGGSGG, GGGGGG, GTGSGT, and 5xGGGGS) forms of the enzyme not only improves its stability, but also increases the mutual mobility of DNA-affinity domains.

ДНК-экзотрансфераза быка (TdT)EcSSBмолекулярная динамикахимерный ферментоцДНК-связывающий белок (SSB)полипептидные линкерыbovine DNA exotransferase (TdT)EcSSBmolecular dynamicsfusion enzymeDNA binding protein (SSB)polypeptide linkersРабота выполнена при поддержке гранта Белорусского республиканского фонда фундаментальных исследований Х21М-056 и задания 20210422 Государственной программы научных исследований «Химические процессы, реагенты и технологии, биорегуляторы и биооргхимия» на 2021–2025 годы.The work is carried out with the support of the grant of the Belarusian Republican Foundation for Fundamental Research Х21М-056 and the task 20210422 of the State Program of Scientific Research “Chemical pro- cesses, reagents and technologies, bioregulators and bioorgc- hemistry” for 2021–2025.ReferencesFowler, J. D. Biochemical, Structural, and Physiological Characterization of Terminal Deoxynucleotidyl Transferase / J. D. Fowler, Z. Suo // Chem. 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The authors declare that there are no conflicts of interest present.