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Characterization and Recombinant Expression of Terebrid Venom Peptide from Terebra guttata

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, maki...

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Autores principales: Moon, John, Gorson, Juliette, Wright, Mary Elizabeth, Yee, Laurel, Khawaja, Samer, Shin, Hye Young, Karma, Yasmine, Musunri, Rajeeva Lochan, Yun, Michelle, Holford, Mande
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4810208/
https://www.ncbi.nlm.nih.gov/pubmed/26950153
http://dx.doi.org/10.3390/toxins8030063
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author Moon, John
Gorson, Juliette
Wright, Mary Elizabeth
Yee, Laurel
Khawaja, Samer
Shin, Hye Young
Karma, Yasmine
Musunri, Rajeeva Lochan
Yun, Michelle
Holford, Mande
author_facet Moon, John
Gorson, Juliette
Wright, Mary Elizabeth
Yee, Laurel
Khawaja, Samer
Shin, Hye Young
Karma, Yasmine
Musunri, Rajeeva Lochan
Yun, Michelle
Holford, Mande
author_sort Moon, John
collection PubMed
description 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.
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spelling pubmed-48102082016-04-04 Characterization and Recombinant Expression of Terebrid Venom Peptide from Terebra guttata Moon, John Gorson, Juliette Wright, Mary Elizabeth Yee, Laurel Khawaja, Samer Shin, Hye Young Karma, Yasmine Musunri, Rajeeva Lochan Yun, Michelle Holford, Mande Toxins (Basel) Article 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. MDPI 2016-03-03 /pmc/articles/PMC4810208/ /pubmed/26950153 http://dx.doi.org/10.3390/toxins8030063 Text en © 2016 by the authors; licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons by Attribution (CC-BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Moon, John
Gorson, Juliette
Wright, Mary Elizabeth
Yee, Laurel
Khawaja, Samer
Shin, Hye Young
Karma, Yasmine
Musunri, Rajeeva Lochan
Yun, Michelle
Holford, Mande
Characterization and Recombinant Expression of Terebrid Venom Peptide from Terebra guttata
title Characterization and Recombinant Expression of Terebrid Venom Peptide from Terebra guttata
title_full Characterization and Recombinant Expression of Terebrid Venom Peptide from Terebra guttata
title_fullStr Characterization and Recombinant Expression of Terebrid Venom Peptide from Terebra guttata
title_full_unstemmed Characterization and Recombinant Expression of Terebrid Venom Peptide from Terebra guttata
title_short Characterization and Recombinant Expression of Terebrid Venom Peptide from Terebra guttata
title_sort characterization and recombinant expression of terebrid venom peptide from terebra guttata
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4810208/
https://www.ncbi.nlm.nih.gov/pubmed/26950153
http://dx.doi.org/10.3390/toxins8030063
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