Email this article Recombinant production and structure-function study of ts1 toxin from the Brazilian scorpion Tityus serrulatus
- Authors: Shenkarev Z.O.1,2, Shulepko M.A.1,3, Peigneur S.4, Myshkin M.Y.1,2, Berkut A.A.1,2, Vassilevski A.A.1,2, Tytgat J.4, Lyukmanova E.N.1,3, Kirpichnikov M.P.1,3
-
Affiliations:
- Shemyakin & Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences
- Moscow Institute of Physics and Technology
- Lomonosov Moscow State University
- Catholic University of Leuven
- Issue: Vol 484, No 1 (2019)
- Pages: 112-116
- Section: Biochemistry, biophysics, molecular biology
- URL: https://journals.eco-vector.com/0869-5652/article/view/12154
- DOI: https://doi.org/10.31857/S0869-56524841112-116
- ID: 12154
Cite item
Full Text
Abstract
This work introduces an effective bacterial system for the production of β-toxin Ts1, the main component of the Brazilian scorpion Tityus serrulatus venom. Recombinant toxin and its 15N-labeled analogue are obtained by direct expression of the synthetic gene in Escherichia coli, with subsequent folding from the inclusion bodies. NMR spectroscopy data assert that the recombinant toxin is structured in aqueous solution and is composed of a significant fraction of β-structure. Moreover, the formation of a stable Ts1 disulfide-bond isomer of a disordered structure is observed during folding; recombinant Ts1 blocks Na+ current through NaV1.5 channels, without affecting the processes of activation and inactivation. Simultaneously, the effect upon NaV1.4 channels is associated with a shift of the activation curve toward the more negative membrane potentials.
About the authors
Z. O. Shenkarev
Shemyakin & Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences; Moscow Institute of Physics and Technology
Author for correspondence.
Email: zakhar-shenkarev@yandex.ru
Russian Federation, 16/10, Miklukho-Maklaya Street, Moscow, 117997; 9, Institutskij, Dolgoprudny, Moscow region, 141701
M. A. Shulepko
Shemyakin & Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences; Lomonosov Moscow State University
Email: zakhar-shenkarev@yandex.ru
Russian Federation, 16/10, Miklukho-Maklaya Street, Moscow, 117997; 1, Leninskie gory, Moscow, 119991
S. Peigneur
Catholic University of Leuven
Email: zakhar-shenkarev@yandex.ru
Belgium, Leuven
M. Yu. Myshkin
Shemyakin & Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences; Moscow Institute of Physics and Technology
Email: zakhar-shenkarev@yandex.ru
Russian Federation, 16/10, Miklukho-Maklaya Street, Moscow, 117997; 9, Institutskij, Dolgoprudny, Moscow region, 141701
A. A. Berkut
Shemyakin & Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences; Moscow Institute of Physics and Technology
Email: zakhar-shenkarev@yandex.ru
Russian Federation, 16/10, Miklukho-Maklaya Street, Moscow, 117997; 9, Institutskij, Dolgoprudny, Moscow region, 141701
A. A. Vassilevski
Shemyakin & Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences; Moscow Institute of Physics and Technology
Email: zakhar-shenkarev@yandex.ru
Russian Federation, 16/10, Miklukho-Maklaya Street, Moscow, 117997; 9, Institutskij, Dolgoprudny, Moscow region, 141701
J. Tytgat
Catholic University of Leuven
Email: zakhar-shenkarev@yandex.ru
Belgium, Leuven
E. N. Lyukmanova
Shemyakin & Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences; Lomonosov Moscow State University
Email: zakhar-shenkarev@yandex.ru
Russian Federation, 16/10, Miklukho-Maklaya Street, Moscow, 117997; 1, Leninskie gory, Moscow, 119991
M. P. Kirpichnikov
Shemyakin & Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences; Lomonosov Moscow State University
Email: zakhar-shenkarev@yandex.ru
Academician of the RAS
Russian Federation, 16/10, Miklukho-Maklaya Street, Moscow, 117997; 1, Leninskie gory, Moscow, 119991References
- Catterall W.A. // J. Physiol. 2012. V. 590. P. 2577–2589.
- Stevens M., Peigneur S., Tytgat J. // Front. Pharmacol. 2011. V. 2. P. 71.
- Mӓnnikkӧ R., Shenkarev Z.O., Thor M.G., Berkut A.A., Myshkin M. Y., Paramonov A.S., Kulbatskii D.S., Kuzmin D.A., Sampedro Castaneda M., King L., Wilson E.R., Lyukmanova E.N., Kirpichnikov M.P., Schorge S., Bosmans F., Hanna M.G., Kullmann D. M., Vassilevski A. A. // Proc. Nat. Acad. Sci. USA. 2018. V. 115. P. 4495–4500.
- Pinheiro C.B., Marangoni S., Toyama M.H. et al. // Acta Crystallogr. D Biol. Crystallogr. 2003. V. 59. P. 405–415.
- Barhanin J., Giglio J.R., Leopold P., et al. // J. Biol. Chem. 1982. V. 257. P. 12553–12558.
- Martin-Eauclaire M.F., Bougis P.E., de Lima M.E. // Toxicon. 2018. V. 152. P. 106–120.
- Bosmans F., Martin-Eauclaire M.F., Tytgat J. // Toxicol. Appl. Pharmacol. 2007. V. 218. P. 45–51.
- Peigneur S., Cologna C.T., Cremonez C.M., et al. // Neuropharmacology. 2015. V. 95. P. 269–277.
- Bosmans F., Martin-Eauclaire M.F., Swartz K.J. // Nature 2008. V. 456. P. 202–208.
- Loret E.P., Sampieri F., Roussel A., et al. // Proteins: Struct., Funct., Bioinf. 1990. V. 8. P. 164–172.
- Dang B., Kubota T., Correa A.M., et al. // Angew. Chem. Int. Ed. 2014. V. 53. P. 8970–8974.
- Coelho V.A., Cremonez C.M., Anjolette F.A., et al. // Toxicon. 2014. V. 83. P. 15–21.
- Люкманова Е.Н., Шулепко М.А., Шенкарев З.О., Долгих Д.А., Кирпичников М.П. // Биоорг. химия. 2010. Т. 36. № 2. С. 149–158.
- Pluzhnikov K., Vassilevski A., Korolkova Y., Fisyunov A., Iegorova O., Krishtal O, Grishin E. // Toxicon. 2007. V. 50. № 7. P. 993–1004.
- Shulepko M.A., Lyukmanova E.N., Shenkarev Z.O., Dubovskii P.V., Astapova M.V., Feofanov A.V., Arseniev A.S., Utkin Y.N., Kirpichnikov M.P., Dolgikh D.A. // Protein Expr. Purif. 2017. V. 130. P. 13–20.
Supplementary files
