Characterization and Recombinant Expression of Terebrid Venom Peptide from Terebra guttata
AbstractVenom peptides found in terebrid snails expand the toolbox of active compounds that can be applied to investigate cellular physiology and can be further developed as future therapeutics. However, unlike other predatory organisms, such as snakes, terebrids produce very small quantities of venom, making it difficult to obtain sufficient amounts for biochemical characterization. Here, we describe the first recombinant expression and characterization of terebrid peptide, teretoxin Tgu6.1, from Terebra guttata. Tgu6.1 is a novel forty-four amino acid teretoxin peptide with a VI/VII cysteine framework (C–C–CC–C–C) similar to O, M and I conotoxin superfamilies. A ligation-independent cloning strategy with an ompT protease deficient strain of E. coli was employed to recombinantly produce Tgu6.1. Thioredoxin was introduced in the plasmid to combat disulfide folding and solubility issues. Specifically Histidine-6 tag and Ni-NTA affinity chromatography were applied as a purification method, and enterokinase was used as a specific cleavage protease to effectively produce high yields of folded Tgu6.1 without extra residues to the primary sequence. The recombinantly-expressed Tgu6.1 peptide was bioactive, displaying a paralytic effect when injected into a Nereis virens polychaete bioassay. The recombinant strategy described to express Tgu6.1 can be applied to produce high yields of other disulfide-rich peptides. View Full-Text
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Moon, J.; Gorson, J.; Wright, M.E.; Yee, L.; Khawaja, S.; Shin, H.Y.; Karma, Y.; Musunri, R.L.; Yun, M.; Holford, M. Characterization and Recombinant Expression of Terebrid Venom Peptide from Terebra guttata. Toxins 2016, 8, 63.
Moon J, Gorson J, Wright ME, Yee L, Khawaja S, Shin HY, Karma Y, Musunri RL, Yun M, Holford M. Characterization and Recombinant Expression of Terebrid Venom Peptide from Terebra guttata. Toxins. 2016; 8(3):63.Chicago/Turabian Style
Moon, John; Gorson, Juliette; Wright, Mary E.; Yee, Laurel; Khawaja, Samer; Shin, Hye Y.; Karma, Yasmine; Musunri, Rajeeva L.; Yun, Michelle; Holford, Mande. 2016. "Characterization and Recombinant Expression of Terebrid Venom Peptide from Terebra guttata." Toxins 8, no. 3: 63.