Cyclotide isolation and characterization

Craik, David J., Henriques, Sonia Troeira, Mylne, Joshua S. and Wang, Conan K. (2012). Cyclotide isolation and characterization. In David A. Hopwood (Ed.), Natural product biosynthesis by microorganisms and plants, part B (pp. 37-62) Burlington, USA: Elsevier. doi:10.1016/B978-0-12-394291-3.00024-1

Author Craik, David J.
Henriques, Sonia Troeira
Mylne, Joshua S.
Wang, Conan K.
Title of chapter Cyclotide isolation and characterization
Title of book Natural product biosynthesis by microorganisms and plants, part B
Place of Publication Burlington, USA
Publisher Elsevier
Publication Year 2012
Sub-type Research book chapter (original research)
DOI 10.1016/B978-0-12-394291-3.00024-1
Series Methods in enzymology
ISBN 9780123942913
ISSN 0076-6879
Editor David A. Hopwood
Volume number 516
Chapter number 3
Start page 37
End page 62
Total pages 26
Total chapters 17
Collection year 2013
Language eng
Abstract/Summary Cyclotides are disulfide-rich cyclic peptides produced by plants with the presumed natural function of defense agents against insect pests. They are present in a wide range of plant tissues, being ribosomally synthesized via precursor proteins that are posttranslationally processed to produce mature peptides with a characteristic cyclic backbone and cystine knot motif associated with their six conserved cysteine residues. Their processing is not fully understood but involves asparaginyl endoproteinase activity. In addition to interest in their defense roles and their unique topologies, cyclotides have attracted attention as potential templates in peptide-based drug design applications. This chapter provides protocols for the isolation of cyclotides from plants, their detection and sequencing by mass spectrometry, and their structural analysis by NMR, as well as describing methods for the isolation of nucleic acid sequences that encode their precursor proteins. Assays to assess their membrane-binding interactions are also described. These protocols provide a “starter kit” for researchers entering the cyclotide field.
Keyword Cyclic peptides
Kalata B1
Mass spectometry
Membrane binding
Q-Index Code BX
Q-Index Status Confirmed Code
Institutional Status UQ

Document type: Book Chapter
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Created: Thu, 17 Jan 2013, 15:21:10 EST by Susan Allen on behalf of Institute for Molecular Bioscience