title: A New, High Yield, Rapid, and Cost-Effective Protocol to Deprotection of Cysteine-Rich Conopeptide, Omega-Conotoxin MVIIA.
abstract: Almost all conopeptides purified from Conus venoms are cysteine-rich peptides. Among them, omega-conotoxin MVIIA, FDA approved peptide drug (Prialt(®)), selected as a cysteine-rich model that its protection from oxidation is critical during solid phase synthesis. Deprotection of cysteines is a crucial step after peptide synthesis. The current study aimed to set up a new highly efficient deprotection protocol for omega-conotoxin MVIIA. Deprotection was performed based on mercury acetate with significant major modification. The protocol accomplished based on the best molar ratio of peptide/mercury/2-ME that adjusted to 0.2 mm/3 mm/10 mm (50 μg/1 mg/10 μL). The yield and purity of omega-conotoxin MVIIA obtained at 93 and 95%, respectively. The total time of protocol shortened to 90 min instead of 6-20 h in routine methods. In this study, omega-conotoxin MVIIA was recovered in high yield and in the shortest time. Despite of other known protocols, molar ratio adjusted to minimum amount. In conclusion, this protocol would be suggested to cost-effective deprotection of thiol groups for similar cysteine-rich peptides.
authors.affiliation: Department of Marine Biology, Faculty of Marine Sciences and Technologies, Science and Research Branch, Islamic Azad University, Tehran, Iran.
authors.forename: Seyed Sahand
authors.initials: SS
authors.lastname: Eisapoor
authors.affiliation: Department of Marine Biology, Faculty of Marine Sciences and Technologies, Science and Research Branch, Islamic Azad University, Tehran, Iran.
authors.forename: Shahla
authors.initials: S
authors.lastname: Jamili
authors.affiliation: Biotechnology Research Center, Medical Biotechnology Department, Venom and Biotherapeutics Molecules Lab, Pasteur Institute of Iran, Tehran, Iran.
authors.forename: Delavar
authors.initials: D
authors.lastname: Shahbazzadeh
authors.affiliation: Department of Marine Biology, Faculty of Marine Sciences and Technologies, Science and Research Branch, Islamic Azad University, Tehran, Iran.
authors.forename: Pargol
authors.initials: P
authors.lastname: Ghavam Mostafavi
authors.affiliation: Biotechnology Research Center, Medical Biotechnology Department, Venom and Biotherapeutics Molecules Lab, Pasteur Institute of Iran, Tehran, Iran.
authors.forename: Kamran
authors.initials: K
authors.lastname: Pooshang Bagheri
mesh_terms: D002851:Chromatography, High Pressure Liquid; D003362:Cost-Benefit Analysis; D003545:Cysteine / Q000032:analysis*; D004591:Electrophoresis, Polyacrylamide Gel; D013058:Mass Spectrometry; D020960:omega-Conotoxins / Q000737:chemistry*
chemical_list: D020960:omega-Conotoxins; C078452:ziconotide; D003545:Cysteine
keywords: cysteine-rich peptides; deprotection; marine cone snails; omega-conotoxin MVIIA