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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| Format: | Article |
| Language: | English |
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MDPI AG
2016-03-01
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| Series: | Toxins |
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| Online Access: | http://www.mdpi.com/2072-6651/8/3/63 |
| _version_ | 1811186289584635904 |
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| author | John Moon Juliette Gorson Mary Elizabeth Wright Laurel Yee Samer Khawaja Hye Young Shin Yasmine Karma Rajeeva Lochan Musunri Michelle Yun Mande Holford |
| author_facet | John Moon Juliette Gorson Mary Elizabeth Wright Laurel Yee Samer Khawaja Hye Young Shin Yasmine Karma Rajeeva Lochan Musunri Michelle Yun Mande Holford |
| author_sort | John Moon |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-04-11T13:43:05Z |
| format | Article |
| id | doaj.art-7df2cd69cd054d89b59d2b7e8af2e7d7 |
| institution | Directory Open Access Journal |
| issn | 2072-6651 |
| language | English |
| last_indexed | 2024-04-11T13:43:05Z |
| publishDate | 2016-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Toxins |
| spelling | doaj.art-7df2cd69cd054d89b59d2b7e8af2e7d72022-12-22T04:21:10ZengMDPI AGToxins2072-66512016-03-01836310.3390/toxins8030063toxins8030063Characterization and Recombinant Expression of Terebrid Venom Peptide from Terebra guttataJohn Moon0Juliette Gorson1Mary Elizabeth Wright2Laurel Yee3Samer Khawaja4Hye Young Shin5Yasmine Karma6Rajeeva Lochan Musunri7Michelle Yun8Mande Holford9Hunter College, City University of New York, Belfer Research Center 413 E. 69th Street, New York, NY 10021, USAHunter College, City University of New York, Belfer Research Center 413 E. 69th Street, New York, NY 10021, USAHunter College, City University of New York, Belfer Research Center 413 E. 69th Street, New York, NY 10021, USAHunter College, City University of New York, Belfer Research Center 413 E. 69th Street, New York, NY 10021, USAHunter College, City University of New York, Belfer Research Center 413 E. 69th Street, New York, NY 10021, USAHunter College, City University of New York, Belfer Research Center 413 E. 69th Street, New York, NY 10021, USAHunter College, City University of New York, Belfer Research Center 413 E. 69th Street, New York, NY 10021, USANew York Genome Center, 101 Avenue of the Americas, New York, NY 10013, USAHunter College, City University of New York, Belfer Research Center 413 E. 69th Street, New York, NY 10021, USAHunter College, City University of New York, Belfer Research Center 413 E. 69th Street, New York, NY 10021, USAVenom 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.http://www.mdpi.com/2072-6651/8/3/63Terebridaevenom peptidesteretoxinsrecombinant synthesispolychaete assaydisulfide-rich peptidesConoideasnail venom |
| spellingShingle | John Moon Juliette Gorson Mary Elizabeth Wright Laurel Yee Samer Khawaja Hye Young Shin Yasmine Karma Rajeeva Lochan Musunri Michelle Yun Mande Holford Characterization and Recombinant Expression of Terebrid Venom Peptide from Terebra guttata Toxins Terebridae venom peptides teretoxins recombinant synthesis polychaete assay disulfide-rich peptides Conoidea snail venom |
| 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 | Terebridae venom peptides teretoxins recombinant synthesis polychaete assay disulfide-rich peptides Conoidea snail venom |
| url | http://www.mdpi.com/2072-6651/8/3/63 |
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