The structure of a two-disulfide intermediate assists in elucidating the oxidative folding pathway of a cyclic cystine knot protein.
We have determined the three-dimensional structure of a two-disulfide intermediate (Cys(8)-Cys(20), Cys(14)-Cys(26)) on the oxidative folding pathway of the cyclotide MCoTI-II. Cyclotides have a range of bioactivities and, because of their exceptional stability, have been proposed as potential molecular scaffolds for drug design applications. The three-dimensional structure of the stable two-disulfide intermediate shows for the most part identical secondary and tertiary structure to the native state. The only exception is a flexible loop, which is collapsed onto the protein core in the native state, whereas in the intermediate it is more loosely associated with the remainder of the protein. The results suggest that the native fold of the peptide does not represent the free energy minimum in the absence of the Cys(1)-Cys(18) disulfide bridge and that although there is not a large energy barrier, the peptide must transiently adopt an energetically unfavorable state before the final disulfide can form.
Cemazar
Masa
M
Institute for Molecular Bioscience, University of Queensland, Brisbane, QLD 4072, Australia.
Joshi
Ajinkya
A
Daly
Norelle L
NL
Mark
Alan E
AE
Craik
David J
DJ
D003198:Computer Simulation; D047169:Cyclotides / Q000737:chemistry*; D047168:Cystine Knot Motifs; D004220:Disulfides / Q000737:chemistry; D007536:Isomerism; D008958:Models, Molecular; D019906:Nuclear Magnetic Resonance, Biomolecular; D010084:Oxidation-Reduction; D010940:Plant Proteins / Q000737:chemistry; D017510:Protein Folding
D047169:Cyclotides; D004220:Disulfides; D010940:Plant Proteins