vestnik-bio-msuВестник Московского университета. Серия 16. БиологияVestnik Moskovskogo universiteta. Seriya 16. Biologiya0137-0952Lomonosov Moscow State University, School of Biologyvestnik-bio-msu-1134Research ArticleОРИГИНАЛЬНОЕ ИССЛЕДОВАНИЕRESEARCH ARTICLEРекомбинантные пептиды Ce1 и Ce4 из яда скорпиона Centruroides elegans и их взаимодействие с гибридными каналами KcsA-K V 1.X (x = 1, 3, 6)Recombinant peptides Ce1 and Ce4 from venom of scorpion Centruroides elegans and their interactions with hybrid channels KcsA-K V 1.x (x = 1, 3, 6)https://orcid.org/0000-0003-0693-7091ОрловН. А.OrlovN. A.

 аспирант, вед. инженер, инженер лаборатории оптической микроскопии и спектроскопии биомолекул

Тел. 8-495-336-64-55

 Россия, 119234, г. Москва, Ленинские горы, д. 1, стр. 12 

 Россия, 117997, г. Москва, ул. Миклухо-Маклая, д. 16/10 

 518172, China 

 1–12 Leninskie gory, Moscow, 119234, Russia 

 16/10 Miklukho-Maklaya str., Moscow, 117997, Russia 

 518172, China 

n.orlov858@yandex.ruhttps://orcid.org/0000-0002-1586-3349ЯкимовС. А.YakimovS. A.

 инженер-исследователь группы нанобиоинженерии

Тел. 8-495-330-66-38

 Россия, 117997, г. Москва, ул. Миклухо-Маклая, д. 16/10 

 16/10 Miklukho-Maklaya str., Moscow, 117997, Russia 

sa-yakimov@yandex.ruhttps://orcid.org/0000-0002-7216-1618НекрасоваО. В.NekrasovaO. V.

 канд. биол. наук, руководитель группы нанобиоинженерии отдела биоинженерии

Тел. 8-495-330-66-38

 Россия, 117997, г. Москва, ул. Миклухо-Маклая, д. 16/10 

 16/10 Miklukho-Maklaya str., Moscow, 117997, Russia 

okatja@yandex.ruhttps://orcid.org/0000-0002-1596-9506ФеофановА. В.FeofanovA. V.

докт. биол. наук, проф., руководитель лаборатории оптической микроскопии и спектроскопии биомолекул

Тел. 8-495-336-64-55

 Россия, 119234, г. Москва, Ленинские горы, д. 1, стр. 12 

 Россия, 117997, г. Москва, ул. Миклухо-Маклая, д. 16/10 

 1–12 Leninskie gory, Moscow, 119234, Russia 

 16/10 Miklukho-Maklaya str., Moscow, 117997, Russia 

avfeofanov@yandex.ruКафедра биоинженерии, биологический факультет, Московский государственный университет имени М.В. Ломоносова; Институт биоорганической химии имени академиков М.М. Шемякина и Ю.А. Овчинникова РАН; School of Biology, Shenzhen MSU-BIT University, No. 1, International University Park Road, Dayun New Town, Longgang District, Shenzhen, Guangdong ProvinceРоссияDepartment of Bioengineering, School of Biology, Lomonosov Moscow State University; Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences; School of Biology, Shenzhen MSU-BIT University, No. 1, International University Park Road, Dayun New Town, Longgang District, Shenzhen, Guangdong ProvinceRussian FederationИнститут биоорганической химии имени академиков М.М. Шемякина и Ю.А. Овчинникова РАНРоссияShemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of SciencesRussian FederationКафедра биоинженерии, биологический факультет, Московский государственный университет имени М.В. Ломоносова; Институт биоорганической химии имени академиков М.М. Шемякина и Ю.А. Овчинникова РАН;РоссияDepartment of Bioengineering, School of Biology, Lomonosov Moscow State University; Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of SciencesRussian Federation202215062022772122–129Copyright © Орлов Н.А., Якимов С.А., Некрасова О.В., Феофанов А.В., 20222022Орлов Н.А., Якимов С.А., Некрасова О.В., Феофанов А.В.Orlov N.A., Yakimov S.A., Nekrasova O.V., Feofanov A.V.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://vestnik-bio-msu.elpub.ru/jour/article/view/1134

Разработана методика получения рекомбинантных функционально-активных пептидов Ce1 и Ce4 из яда скорпиона Centruroides elegans в системе экспрессии Escherichia coli. Выходы Ce1 и Ce4 составили соответственно 6,5 и 12 мг с литра культуры (плотность клеток около 5 оптических единиц). Свойства полученных пептидов исследованы с применением биоинженерных систем на основе гибридных каналов KcsA-Kv1.x (х = 1, 3, 6), содержащих сайты связывания блокаторов соответствующих эукариотических калиевых каналов семейства Kv1. Показано, что рекомбинантные Ce1 и Ce4 вплоть до микромолярных концентраций не проявляют сродства к сайтам связывания каналов Kv1.1 и Kv1.6 и, подобно природным пептидам, селективно взаимодействуют с сайтом связывания канала Kv1.3: кажущиеся константы диссоциации комплексов KcsA-Kv1.3 с рекомбинантными Ce1 и Ce4 равны, соответственно, 50±10 и 200±30 нМ (среднее ± стандартная ошибка среднего).

A technique has been developed for obtaining recombinant functionally-active peptides Ce1 and Ce4 from the venom of the scorpion Centruroides elegans in the E. coli expression system. The yields of peptides Ce1 and Ce4 were 6.5 and 12 mg per liter of culture, respectively. The properties of the obtained peptides were studied using bioengineered systems based on hybrid KV1 channels KcsA-Kv1.x (x=1, 3, 6) containing blocker binding sites of the corresponding eukaryotic potassium channels of Kv1-family. It has been shown that recombinant Ce1 and Ce4 do not exhibit affinity to the binding sites of Kv1.1 and Kv1.6 channels up to micromolar concentrations and, like natural peptides, selectively interact with the binding site of Kv1.3 channel: the apparent dissociation constants of KcsA-Kv1.3 complexes with recombinant Ce1 and Ce4 are 50±10 and 200±30 nM, respectively.

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