title: Recombinant Expression of Cyclotides Using Split Inteins.
abstract: Cyclotides are fascinating microproteins (≈30 residues long) present in several families of plants that share a unique head-to-tail circular knotted topology of three disulfide bridges, with one disulfide penetrating through a macrocycle formed by the two other disulfides and inter-connecting peptide backbones, forming what is called a cystine knot topology. Naturally occurring cyclotides have shown to posses various pharmacologically relevant activities and have been reported to cross cell membranes. Altogether, these features make the cyclotide scaffold an excellent molecular framework for the design of novel peptide-based therapeutics, making them ideal substrates for molecular grafting of biological peptide epitopes. In this chapter we describe how to express a native folded cyclotide using intein-mediated protein trans-splicing in live Escherichia coli cells.
authors.affiliation: Department of Pharmacology and Pharmaceutical Sciences, University of Southern California, Los Angeles, CA, 90089-9121, USA.
authors.forename: Krishnappa
authors.initials: K
authors.lastname: Jagadish
authors.affiliation: Department of Pharmacology and Pharmaceutical Sciences, University of Southern California, Los Angeles, CA, 90089-9121, USA. jcamarer@usc.edu.|Department of Chemistry, University of Southern California, Los Angeles, CA, 90089-9121, USA. jcamarer@usc.edu.
authors.forename: Julio A
authors.initials: JA
authors.lastname: Camarero
mesh_terms: D047169:Cyclotides / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D047668:Inteins; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification
chemical_list: D047169:Cyclotides; D011993:Recombinant Fusion Proteins
keywords: CCK motif; Cyclotides; Npu intein; Protein trans-splicing; Split-intein