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RAS BiologyБиоорганическая химия Russian Journal of Bioorganic Chemistry

  • ISSN (Print) 0132-3423
  • ISSN (Online) 1998-2860

Three-Finger Viper Toxins – cDNA Cloning and Expression in Using a Chimeric (Hybrid) Construction with a Partner Protein SUMO

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PII
S19982860S0132342325050089-1
DOI
10.7868/S1998286025050089
Publication type
Article
Status
Published
Authors
D.A. Sukhov
  • Affiliation:
    Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences
    MIREA – Russian Technological University
L.O. Ojomoko
  • Affiliation: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences
I.V. Shelukhina
  • Affiliation: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences
M.V. Vladykina
  • Affiliation: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences
V.Yu. Kost
  • Occupation: Department of Chemical and Structural Biology
  • Affiliation: Weizmann Institute of Science
R.Kh. Ziganshin
  • Affiliation: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences
O.V. Geraskina
  • Affiliation: M.V. Lomonosov Moscow State University, Faculty of Biology
S.V. Balandin
  • Affiliation: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences
T.V. Ovchinnikova
  • Affiliation: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences
V.I. Tsetlin
  • Affiliation: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences
Yu.N. Utkin
  • Affiliation: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences
Volume/ Edition
Volume 51 / Issue number 5
Pages
831-843
Abstract
Three-finger toxins (TFTs) form one of the most abundant families of toxins in snake venoms. TFTs are common for most Elapid venoms, but are almost never found in viper venoms. Using the venom glands of the vipers and , 21 cDNA clones encoding this group of toxins were obtained. The amino acid sequences of 9 TFTs were deduced from the obtained cDNA sequences. The identified sequences have signal peptides containing 19-21 amino acid residues, followed by a mature protein consisting of 67 residues. All viper TFTs belong to the group of non-conventional toxins, and their sequences contain 9 cysteine residues. The TFT encoded by one of the transcripts was obtained by heterologous expression in cells as a fusion protein with the plant partner protein SUMO, followed by cleavage with the specific plant protease BdSENP1 and chromatographic purification. The structure of the obtained protein was confirmed by mass spectrometry. Analysis of its biological activity showed that this toxin is a weak antagonist of nicotinic acetylcholine receptors of the neuronal α7 and α3β2 subtypes. Using the fusion protein with SUMO, we also attempted to obtain the TFT Aze-2 of the viper , the amino acid sequence of which was previously established by us as a result of transcriptome analysis of the venom gland of , and the protein itself was identified in minimal quantities in the venom of this snake. However, a toxin exactly corresponding in mass to Aze-2 could not be obtained using this approach. Thus, as a result of the work, the amino acid sequences of 9 viper TFTs were established, one of which was obtained by gene expression in cells and showed the ability to interact with nicotinic acetylcholine receptors of the neuronal α7 and α3β2 subtypes.
Keywords
трехпетельные токсины клонирование экспрессия SUMO масс-спектрометрия никотиновый холинорецептор
Date of publication
01.05.2025
Year of publication
2025
Number of purchasers
0
Views
4

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