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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.

SUBMITTER: Jagadish K 

PROVIDER: S-EPMC5117428 | biostudies-literature | 2017

REPOSITORIES: biostudies-literature

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Recombinant Expression of Cyclotides Using Split Inteins.

Jagadish Krishnappa K   Camarero Julio A JA  

Methods in molecular biology (Clifton, N.J.) 20170101


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 memb  ...[more]

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