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Issue 4, 2012
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Recombinant production of rhesus θ-defensin-1 (RTD-1) using a bacterial expression system

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Abstract

Defensins are antimicrobial peptides that are important in the innate immune defense of mammals. In contrast to mammalian α- and β-defensins, rhesus θ-defensin-1 (RTD-1) comprises only 18 amino acids stabilized by three disulfide bonds and an unusual backbone cyclic topology. In this work we report for the first time the recombinant expression of the fully folded θ-defensin RTD-1 using a bacterial expression system. This was accomplished using an intramolecular native chemical ligation in combination with a modified protein-splicing unit. RTD-1 was produced either in vitro or in vivo. In-cell production of RTD-1 was estimated to reach an intracellular concentration of ∼4 μM. Recombinant RTD-1 was shown to be correctly folded as characterized by heteronuclear-NMR and by its ability to specifically inhibit lethal factor protease. The recombinant production of folded θ-defensins opens the possibility to produce peptide libraries based on this peptide scaffold that could be used to develop in-cell screening and directed evolution technologies.

Graphical abstract: Recombinant production of rhesus θ-defensin-1 (RTD-1) using a bacterial expression system

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Publication details

The article was received on 01 Nov 2011, accepted on 19 Jan 2012 and first published on 10 Feb 2012


Article type: Paper
DOI: 10.1039/C2MB05451E
Citation: Mol. BioSyst., 2012,8, 1359-1365
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    Recombinant production of rhesus θ-defensin-1 (RTD-1) using a bacterial expression system

    A. Gould, Y. Li, S. Majumder, A. E. Garcia, P. Carlsson, A. Shekhtman and J. A. Camarero, Mol. BioSyst., 2012, 8, 1359
    DOI: 10.1039/C2MB05451E

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