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)
Abstract
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.
Keywords
potential-dependent potassium channels,
Kv1,
hybrid potassium channel KcsA-Kv1.3,
laser confocal microscopy,
peptide blockers,
affinity
Supporting agencies: The research was funded by Russian Science Foundation, project number 19-74-30014. The research was partially performed using facilities of the Interdisciplinary Scientific and Educational School of Moscow University “Molecular Technologies of the Living Systems and Synthetic Biology”.
About the Authors
N. A. Orlov
Department 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 Province
Russian Federation
Russian Federation
1–12 Leninskie gory, Moscow, 119234, Russia
16/10 Miklukho-Maklaya str., Moscow, 117997, Russia
518172, China
S. A. Yakimov
Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences
Russian Federation
Russian Federation
16/10 Miklukho-Maklaya str., Moscow, 117997, Russia
O. V. Nekrasova
Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences
Russian Federation
Russian Federation
16/10 Miklukho-Maklaya str., Moscow, 117997, Russia
A. V. Feofanov
Department of Bioengineering, School of Biology, Lomonosov Moscow State University;
Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences
Russian Federation
Russian Federation
1–12 Leninskie gory, Moscow, 119234, Russia
16/10 Miklukho-Maklaya str., Moscow, 117997, Russia
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Review
For citations:
Orlov N.A., Yakimov S.A., Nekrasova O.V., Feofanov A.V. 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). Vestnik Moskovskogo universiteta. Seriya 16. Biologiya. 2022;77(2):122–129. (In Russ.)
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