title: ÐÑÐ´ÐµÐ»ÐµÐ½Ð¸Ðµ ÑÑÐ½ÐºÑÐ¸Ð¾Ð½Ð°Ð»ÑÐ½Ð¾ Ð°ÐºÑÐ¸Ð²Ð½ÑÑ Ð±ÐµÐ»ÐºÐ¾Ð² 3CL-Pro Ð¸ RdRp Ð¸Ð· SARS-CoV-2 Ð´Ð»Ñ in vitro Ð°Ð½Ð°Ð»Ð¸Ð·Ð° Ð¸Ñ Ð²Ð·Ð°Ð¸Ð¼Ð¾Ð´ÐµÐ¹ÑÑÐ²Ð¸Ñ Ñ Ð¿Ð¾ÑÐµÐ½ÑÐ¸Ð°Ð»ÑÐ½ÑÐ¼Ð¸ Ð¸Ð½Ð³Ð¸Ð±Ð¸ÑÐ¾ÑÐ°Ð¼Ð¸
title: Ð²ÑÐ¿ÑÑÐºÐ½Ð°Ñ ÐºÐ²Ð°Ð»Ð¸ÑÐ¸ÐºÐ°ÑÐ¸Ð¾Ð½Ð½Ð°Ñ ÑÐ°Ð±Ð¾ÑÐ° Ð±Ð°ÐºÐ°Ð»Ð°Ð²ÑÐ°
titleType: Subtitle
title: Purification of functionally active 3CL-Pro and RdRp proteins from SARS-Cov-2 for in vitro analysis of their interaction with potential inhibitors
Creators: name: ÐÐ¸Ð´Ð¶Ð¸ÐµÐ²Ð°, ÐÐµÐ´Ð¸Ð½Ð°
Publisher: name: Ð¡Ð°Ð½ÐºÑ-ÐÐµÑÐµÑÐ±ÑÑÐ³ÑÐºÐ¸Ð¹ Ð¿Ð¾Ð»Ð¸ÑÐµÑÐ½Ð¸ÑÐµÑÐºÐ¸Ð¹ ÑÐ½Ð¸Ð²ÐµÑÑÐ¸ÑÐµÑ ÐÐµÑÑÐ° ÐÐµÐ»Ð¸ÐºÐ¾Ð³Ð¾
Year: 2022
Subjects: SARS-CoV-2
Ð¿Ð¾Ð»Ð¸Ð¼ÐµÑÐ°Ð·Ð°
Ð¿ÑÐ¾ÑÐµÐ°Ð·Ð°
Ð¸Ð½Ð³Ð¸Ð±Ð¸ÑÐ¾Ñ
Ñ ÑÐ¾Ð¼Ð°ÑÐ¾Ð³ÑÐ°ÑÐ¸Ñ
polymerase
protease
inhibitor
chromatography

description: ÐÐ°Ð½Ð½Ð°Ñ ÑÐ°Ð±Ð¾ÑÐ° Ð¿Ð¾ÑÐ²ÑÑÐµÐ½Ð° ÑÐ°Ð·ÑÐ°Ð±Ð¾ÑÐºÐµ Ð¿ÑÐ¾ÑÐ¾ÐºÐ¾Ð»Ð° Ð²ÑÐ´ÐµÐ»ÐµÐ½Ð¸Ñ ÑÑÐ½ÐºÑÐ¸Ð¾Ð½Ð°Ð»ÑÐ½Ð¾ Ð°ÐºÑÐ¸Ð²Ð½ÑÑ Ð½ÐµÑÑÑÑÐºÑÑÑÐ½ÑÑ Ð±ÐµÐ»ÐºÐ¾Ð² 3CL-ProÂ (Nsp5) Ð¸Â RdRpÂ (Nsp12) Ð¸Ð·Â SARS-CoV-2, Ð° ÑÐ°ÐºÐ¶Ðµ Ð¸ÑÑÐ»ÐµÐ´Ð¾Ð²Ð°Ð½Ð¸Ñ Ð¸Ñ Ð²Ð·Ð°Ð¸Ð¼Ð¾Ð´ÐµÐ¹ÑÑÐ²Ð¸Ñ Ñ Ð¿Ð¾ÑÐµÐ½ÑÐ¸Ð°Ð»ÑÐ½ÑÐ¼Ð¸ Ð¸Ð½Ð³Ð¸Ð±Ð¸ÑÐ¾ÑÐ°Ð¼Ð¸, Ð¿Ð¾Ð´Ð¾Ð±ÑÐ°Ð½Ð½ÑÐ¼Ð¸ Ð¸Ð· ÑÐ¸ÑÐ»Ð° Ð·Ð°ÑÐµÐ³Ð¸ÑÑÑÐ¸ÑÐ¾Ð²Ð°Ð½Ð½ÑÑ ÑÐµÑÐ°Ð¿ÐµÐ²ÑÐ¸ÑÐµÑÐºÐ¸Ñ Ð¿ÑÐµÐ¿Ð°ÑÐ°ÑÐ¾Ð².Â ÐÐ»Ñ Ð¾ÑÐ¸ÑÑÐºÐ¸ Ð±ÐµÐ»ÐºÐ¾Ð² Ð¸ÑÐ¿Ð¾Ð»ÑÐ·Ð¾Ð²Ð°Ð»Ð¸ÑÑ ÑÐ°ÐºÐ¸Ðµ Ð¼ÐµÑÐ¾Ð´Ñ, ÐºÐ°Ðº Ð¼ÐµÑÐ°Ð»Ð»Ð¾Ð°ÑÑÐ¸Ð½Ð½Ð°Ñ ÑÑÐ¾Ð¼Ð°ÑÐ¾Ð³ÑÐ°ÑÐ¸Ñ, Ð¸Ð¾Ð½Ð¾Ð¾Ð±Ð¼ÐµÐ½Ð½Ð°Ñ ÑÑÐ¾Ð¼Ð°ÑÐ¾Ð³ÑÐ°ÑÐ¸Ñ, Ð¾Ð±ÑÐ°ÑÐ½Ð°Ñ ÑÑÐ¾Ð¼Ð°ÑÐ¾Ð³ÑÐ°ÑÐ¸Ñ Ð½Ð° Ð³ÐµÐ¿Ð°ÑÐ¸Ð½Ðµ, Ð³ÐµÐ»Ñ-ÑÐ¸Ð»ÑÑÑÐ°ÑÐ¸Ñ.Â ÐÑÐ»Ð° Ð¿Ð¾Ð´ÑÐ²ÐµÑÐ¶Ð´ÐµÐ½Ð° Ð¿ÑÐ¾ÑÐµÐ¾Ð»Ð¸ÑÐ¸ÑÐµÑÐºÐ°Ñ Ð°ÐºÑÐ¸Ð²Ð½Ð¾ÑÑÑ Ð²ÑÐ´ÐµÐ»ÐµÐ½Ð½Ð¾Ð³Ð¾ 3CL-ProÂ Ð¸ Ð¿ÑÐ¾Ð²ÐµÐ´ÐµÐ½Â invitroÂ Ð°Ð½Ð°Ð»Ð¸Ð· Ð²Ð·Ð°Ð¸Ð¼Ð¾Ð´ÐµÐ¹ÑÑÐ²Ð¸Ñ Ð±ÐµÐ»ÐºÐ° Ñ Ð¿Ð¾ÑÐµÐ½ÑÐ¸Ð°Ð»ÑÐ½ÑÐ¼Ð¸ Ð¸Ð½Ð³Ð¸Ð±Ð¸ÑÐ¾ÑÐ°Ð¼Ð¸.Â ÐÐ»Ñ ÑÐ²ÑÐ·ÑÐ²Ð°Ð½Ð¸ÑÂ RdRpÂ Ñ ÑÐ°ÐºÑÐ¾ÑÐ°Ð¼Ð¸ Ð¿ÑÐ¾ÑÐµÑÑÐ¸Ð²Ð½Ð¾ÑÑÐ¸Â Nsp7 Ð¸Â Nsp8 Ð±ÑÐ» Ð¿ÑÐ¸Ð³Ð¾ÑÐ¾Ð²Ð»ÐµÐ½ Ð´Ð²ÑÑÐµÐ¿Ð¾ÑÐµÑÐ½ÑÐ¹ Ð ÐÐâÑÑÐ±ÑÑÑÐ°Ñ.Â Â ÐÑÐ»Ð° Ð¿ÑÐ¾Ð²ÐµÑÐµÐ½Ð° ÑÐµÑÐ¼ÐµÐ½ÑÐ°ÑÐ¸Ð²Ð½Ð°Ñ Ð°ÐºÑÐ¸Ð²Ð½Ð¾ÑÑÑ Ð±ÐµÐ»ÐºÐ° Ð² ÐºÐ¾Ð¼Ð¿Ð»ÐµÐºÑÐµ Ñ ÐºÐ¾ÑÐ°ÐºÑÐ¾ÑÐ°Ð¼Ð¸ Ð¸ ÑÑÐ±ÑÑÑÐ°ÑÐ¾Ð¼.Â Ð ÐµÐ·ÑÐ»ÑÑÐ°ÑÑ Ð¿ÑÐ¾Ð´ÐµÐ»Ð°Ð½Ð½Ð¾Ð¹ ÑÐ°Ð±Ð¾ÑÑ Ð¼Ð¾Ð³ÑÑ ÑÐ»ÑÐ¶Ð¸ÑÑ Ð¾ÑÐ½Ð¾Ð²Ð¾Ð¹ Ð´Ð»Ñ Ð´Ð°Ð»ÑÐ½ÐµÐ¹ÑÐ¸Ñ Ð¸ÑÑÐ»ÐµÐ´Ð¾Ð²Ð°Ð½Ð¸Ð¹ Ð¿Ð¾ ÑÐ°Ð·ÑÐ°Ð±Ð¾ÑÐºÐµ Ð¼ÐµÑÐ¾Ð´Ð¸Ðº Ð»ÐµÑÐµÐ½Ð¸Ñ Ð¸ Ð¿ÑÐµÐ¿Ð°ÑÐ°ÑÐ¾Ð², Ð¿Ð¾Ð·Ð²Ð¾Ð»ÑÑÑÐ¸Ñ Ð¿ÑÐµÑÐ²Ð°ÑÑ Ð¶Ð¸Ð·Ð½ÐµÐ½Ð½ÑÐ¹ ÑÐ¸ÐºÐ» ÐºÐ¾ÑÐ¾Ð½Ð°Ð²Ð¸ÑÑÑÐ° ÐµÑÐµ Ð´Ð¾ Ð½Ð°ÑÐ°Ð»Ð° ÑÑÐ°Ð¿Ð° ÑÐ±Ð¾ÑÐºÐ¸ Ð½Ð¾Ð²ÑÑ Ð²Ð¸ÑÑÑÐ½ÑÑ ÑÐ°ÑÑÐ¸Ñ Ð² ÐºÐ»ÐµÑÐºÐµ.
description: The given work is devoted to the development of a functionally active non-structural SARS-CoV-2 3CL-Pro (Nsp5) and RdRp (Nsp12) proteins purification protocol, and to the study of their interaction with potential inhibitors selected from registered therapeutic drugs.Â For protein purification were used such methods as metal-affinity chromatography, ion-exchange chromatography, reverse heparin chromatography, and gel-filtration.Â After confirmation of the proteolytic activity of 3CL-Pro, anÂ in vitroÂ analysis of the interaction of the protein with potential inhibitors was carried out.Â To bind RdRp to Nsp7 and Nsp8 co-factors a double-stranded RNA substrate was prepared. The enzymatic activity of the protein in combination with cofactors and substrate was tested.Â The results of this work can be used for further research on the development of treatment methods and drugs that can interrupt the life cycle of coronavirus even before the assembly of new viral particles in the cell begins.