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

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.