Recombinant Expression of Cyclotides Using Expressed Protein Ligation
| Autor: | Jingtan Su, Julio A. Camarero, Maria Jose Campbell |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Protein Folding Recombinant Fusion Proteins Peptide Cyclotides 010402 general chemistry Protein Engineering 01 natural sciences Epitope Chromatography Affinity Article Inteins 03 medical and health sciences Protein splicing Escherichia coli Disulfides Cloning Molecular Chromatography High Pressure Liquid Plant Proteins chemistry.chemical_classification Oxidative folding Cystine knot Chromatography Agarose 0104 chemical sciences Cyclotide 030104 developmental biology chemistry Biochemistry Cyclization Cystine Electrophoresis Polyacrylamide Gel Cystine Knot Motifs Intein Protein Processing Post-Translational |
| Zdroj: | Methods Mol Biol Expressed Protein Ligation ISBN: 9781071604335 |
| ISSN: | 1940-6029 |
| Popis: | Cyclotides are naturally occurring microproteins (≈30 residues long) present in several families of plants. All cyclotides share a unique head-to-tail circular knotted topology containing three disulfide bridges forming a cystine knot topology. Cyclotides possess high stability to chemical, physical, and biological degradation and have been reported to cross cellular membranes. In addition, naturally occurring and engineered cyclotides have shown to possess various pharmacologically relevant activities. These unique features make the cyclotide scaffold an excellent tool for the design of novel peptide-based therapeutics by using molecular evolution and/or peptide epitope grafting techniques. In this chapter, we provide protocols to recombinantly produce a natively folded cyclotide making use of a standard bacterial expression system in combination with an intein-mediated backbone cyclization with concomitant oxidative folding. |
| Databáze: | OpenAIRE |
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