Identification and characterization of a novel O-superfamily conotoxin from Conus litteratus

A novel conotoxin named lt6c, an O-superfamily conotoxin, was identified from the cDNA library of venom duct of Conus litteratus. The full-length cDNA contains an open reading frame encoding a predicted 22-residue signal peptide, a 22-residue proregion and a mature peptide of 28 amino acids. The signal peptide sequence of lt6c is highly conserved in O-superfamily conotoxins and the mature peptide consists of six cysteines arranged in the pattern of C-C-CC-C-C that is defined the O-superfamily of conotoxins. The mature peptide fused with thioredoxin, 6-His tag, and a Factor Xa cleavage site was successfully expressed in Escherichia coli. About 12 mg lt6c was purified from 1L culture. Under whole-cell patch-clamp mode, lt6c inhibited sodium currents on adult rat dorsal root ganglion neurons. Therefore, lt6c is a novel O-superfamily conotoxin that is able to block sodium channels.