Bioengineering of a Novel Peptide Sequence from the Venom of Conus obscurus

Abstract

The marine cone snail produces one of the fastest prey strikes in the animal kingdom with efficacious venom injection causing prey paralysis and death within seconds. Each snail produces hundreds of conotoxins and has been the source behind discovering and utilizing novel analgesic peptide therapeutics. In this study, we discover, isolate, and synthesize two α3/5 conotoxins derived from the milked venom of Conus obscurus: one novel (α-conotoxin ObI) and one previously found in the venom of Conus striatus (α-SI). We then generate five synthetic analogs, accompanying single and double mutations from the native α-conotoxin ObI. We integrate three post-translational modifications (PTMs) within analog development: N-terminal truncation, proline hydroxylation, and tryptophan bromination. α-Conotoxin ObI demonstrates nanomolar potency towards Poecilia reticulata (LD50) and the Homo sapiens muscle-type nAChR (EC50). Moreover, the analog α-ObI [P9K] displayed enhanced potency in both animal bioassays. The exhibited successful incorporation of 3 PTMs investigates the boundaries of peptide bioengineering in the generation of novel α-conotoxins.