Functional characterization of two novel scorpion sodium channel toxins from Lychas mucronatus

Abstract

The diverse α-scorpion toxins are invaluable pharmacological tools and potential drugs targeting sodium channels, but the pharmacological profiles of most toxins remains unknown so far. Here, we reported pharmacological activities of two novel α-scorpion toxins LmαTX3 and LmαTX5 from the Lychas mucronatus. Using the expression vector pET-28a, the recombinant LmαTX3 and LmαTX5 were separated by RP-HPLC and identified by MALDI-TOF-MS. Subsequently, sodium channels rNav1.2, mNav1.4, hNav1.5 and hNav1.7 were used for evaluating the pharmacological activities of LmαTX3 and LmαTX5 toxins. The electrophysiological experiments showed that both 10 μM recombinant LmαTX3 and LmαTX5 seriously inhibited the fast inactivation of mNav1.4 and hNav1.5 channels, moderately affected hNav1.7 channel, and hardly modulated rNav1.2 channel. The dose–response experiments further indicated the EC50 values of LmαTX3 were 4.98 ± 0.79 μM for mNav1.4, 1.23 ± 0.31 μM for hNav1.5 and 31.46 ± 2.32 μM for hNav1.7 channels, respectively. Similar pharmacological profiles of recombinant LmαTX5 were also observed, and its EC50 values were 4.53 ± 1.38 μM, 1.03 ± 0.43 μM and 67.62 ± 2.31 μM for mNav1.4, hNav1.5 and hNav1.7, respectively. In addition, the recombinant LmαTX3 from the vector pET-14b had much less effect on the fast inactivation of mNav1.4, hNav1.5 and hNav1.7 channels, which indicated that the expression vector pET-14b likely played a critical role in toxin function. Together, these findings first highlighted that scorpion toxins from L. mucronatus were a new molecular resource of discovering pharmacological probes and prospective drugs targeting sodium channels in the future.