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In vitro production of three-finger neurotoxins from snake venoms, a disulfide rich proteins. Problems and their solutions (Review)

On 50-year anniversary of the Institute of Bioorganic Chemistry

Abstract

α-Neurotoxins from snake venom are highly efficient inhibitors of nicotinic acetylcholine receptors (nAChRs). These small proteins with a β-structural organization attract much interest as a tool for studies of nAChR and as prototypes for the development of new pharmaceuticals for the treatment of diseases of the nervous system. However, in vitro production of “three-finger” neurotoxins is complicated by the presence of four or five spatially closed disulfide bonds in their molecules. The present review contains a description of the most frequently used modern approaches for the E. coli expression of recombinant proteins (direct expression, expression as hybrid proteins, and secretion) with an emphasis placed on the recombinant production of snake α-neurotoxins. The methods of E. coli expression of isotopically labeled neurotoxins are described. The proposed solutions will be of broad interest for the bacterial production of other disulfide rich proteins.

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Abbreviations

GSH and GSSH:

reduced and oxidized forms of glutathione

GST:

glutathione-transferase

IC50 :

inhibition constant, at wich 50% of binding sites of receptor is blocked

nAChR:

nicotine acetylcholine receptor

NTII:

neurotoxin of venom of Naja oxiana

NTII/NTI:

chimeric toxin obtained by introduction onto the end of central loop of neurotoxin II of a fragment of the central loop of neurotoxin I of venom of Naja oxiana with additional disulfide bond

STII:

signal peptide of enterotoxin II of E. coli

TRX:

thioredoxine

WTX:

“weak” toxin of venom of Naja kaouthia

ZZ:

domen—synthetic IgG-binding ZZ-domen of protein A of staphillococci

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Correspondence to E. N. Lyukmanova.

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Original Russian Text © E.N. Lyukmanova, M.A. Shulepko, Z.O. Shenkarev, D.A. Dolgikh, M.P. Kirpichnikov, 2010, published in Bioorganicheskaya Khimiya, 2010, Vol. 36, No. 2, pp. 149–158.

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Lyukmanova, E.N., Shulepko, M.A., Shenkarev, Z.O. et al. In vitro production of three-finger neurotoxins from snake venoms, a disulfide rich proteins. Problems and their solutions (Review). Russ J Bioorg Chem 36, 137–145 (2010). https://doi.org/10.1134/S1068162010020019

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  • DOI: https://doi.org/10.1134/S1068162010020019

Key words

  • α-neurotoxins
  • nicotinic acetylcholine receptor
  • bacterial expression
  • three-finger proteins
  • expression in Escherichia coli