title: Characterization and Recombinant Expression of Terebrid Venom Peptide from Terebra guttata.
abstract: Venom 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.
authors.affiliation: Hunter College, City University of New York, Belfer Research Center 413 E. 69th Street, New York, NY 10021, USA. John.jj.moon@gmail.com.
authors.forename: John
authors.initials: J
authors.lastname: Moon
authors.affiliation: Hunter College, City University of New York, Belfer Research Center 413 E. 69th Street, New York, NY 10021, USA. jgorson@gradcenter.cuny.edu.|The Graduate Center, City University of New York, 365 Fifth Avenue, New York, NY 10016, USA. jgorson@gradcenter.cuny.edu.
authors.forename: Juliette
authors.initials: J
authors.lastname: Gorson
authors.affiliation: Hunter College, City University of New York, Belfer Research Center 413 E. 69th Street, New York, NY 10021, USA. lzwright@gmail.com.|The Graduate Center, City University of New York, 365 Fifth Avenue, New York, NY 10016, USA. lzwright@gmail.com.
authors.forename: Mary Elizabeth
authors.initials: ME
authors.lastname: Wright
authors.affiliation: Hunter College, City University of New York, Belfer Research Center 413 E. 69th Street, New York, NY 10021, USA. yeelaurel93@gmail.com.
authors.forename: Laurel
authors.initials: L
authors.lastname: Yee
authors.affiliation: Hunter College, City University of New York, Belfer Research Center 413 E. 69th Street, New York, NY 10021, USA. samerk28@gmail.com.
authors.forename: Samer
authors.initials: S
authors.lastname: Khawaja
authors.affiliation: Hunter College, City University of New York, Belfer Research Center 413 E. 69th Street, New York, NY 10021, USA. hyshin1209@gmail.com.
authors.forename: Hye Young
authors.initials: HY
authors.lastname: Shin
authors.affiliation: Hunter College, City University of New York, Belfer Research Center 413 E. 69th Street, New York, NY 10021, USA. yrkarma@gmail.com.
authors.forename: Yasmine
authors.initials: Y
authors.lastname: Karma
authors.affiliation: New York Genome Center, 101 Avenue of the Americas, New York, NY 10013, USA. raj.musunuri@gmail.com.
authors.forename: Rajeeva Lochan
authors.initials: RL
authors.lastname: Musunri
authors.affiliation: Hunter College, City University of New York, Belfer Research Center 413 E. 69th Street, New York, NY 10021, USA. yunm11@gmail.com.
authors.forename: Michelle
authors.initials: M
authors.lastname: Yun
authors.affiliation: Hunter College, City University of New York, Belfer Research Center 413 E. 69th Street, New York, NY 10021, USA. mholford@hunter.cuny.edu.|The Graduate Center, City University of New York, 365 Fifth Avenue, New York, NY 10016, USA. mholford@hunter.cuny.edu.|The American Museum of Natural History, 79th Street at Central Park West, New York, NY 10026, USA. mholford@hunter.cuny.edu.
authors.forename: Mande
authors.initials: M
authors.lastname: Holford
mesh_terms: D000818:Animals; D008978:Mollusk Venoms / Q000737:chemistry*; D010243:Paralysis / Q000139:chemically induced; D010455:Peptides / Q000235:genetics* / Q000633:toxicity*; D010957:Plasmids; D011077:Polychaeta / Q000187:drug effects*; D011995:Recombination, Genetic; D012908:Snails
chemical_list: D008978:Mollusk Venoms; D010455:Peptides
keywords: Conoidea; Terebridae; disulfide-rich peptides; polychaete assay; recombinant synthesis; snail venom; teretoxins; venom peptides