Abstract
The cyclic peptide sunflower trypsin inhibitor 1 (SFTI-1) blocks trypsin and is a promising drug lead and protein engineering scaffold. We show that SFTI-1 and the newfound SFT-L1 are buried within PawS1 and PawS2, precursors for seed storage protein albumins. Proalbumins are matured by asparaginyl endopeptidase, which we show is required to liberate both ends of SFTI-1 as well as to mature PawS1 albumin. Thus, these peptides emerge from within an albumin precursor by the action of albumin's own processing enzyme.
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Acknowledgements
The authors thank staff at the Arizona Genomics Institute (University of Arizona) for providing Helianthus cDNA clones, the Compositae Genome Project website (http://cgpdb.ucdavis.edu/) that was supported by the United States Department of Agriculture Initiative for Future Agriculture and Food Systems program and National Science Foundation Plant Genome Program for EST data, S. Dillon (Australian Plant Genetic Resource Information Service) for Helianthus seeds, I. Hara-Nishimura (University of Kyoto) for aep null seeds, A. Argyros for technical assistance, H. Schirra (University of Queensland) for providing the structural image used in Supplementary Figure 1 as well as R. Jung, M. Anderson, D. Ortiz-Barrientos and S. Balasubramanian for comments. D.J.C. is a National Health and Medical Research Council Professorial Fellow. J.S.M. is an Australian Research Council Queen Elizabeth II Fellow (DP0879133) and The John S. Mattick Fellow. N.L.D. is a Queensland Smart State Fellow.
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J.S.M., N.L.D. and D.J.C. conceived the study; J.S.M., A.H.C., A.G.E. and E.J.M. performed the experiments, N.L.D. obtained the SFT-L1 structure; M.L.C. and A.J. performed MS studies; J.S.M., N.L.D. and M.L.C. analyzed the data; J.S.M. and D.J.C. wrote the paper.
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Supplementary Methods, Supplementary Figures 1–13 and Supplementary Tables 1–8 (PDF 6835 kb)
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Mylne, J., Colgrave, M., Daly, N. et al. Albumins and their processing machinery are hijacked for cyclic peptides in sunflower. Nat Chem Biol 7, 257–259 (2011). https://doi.org/10.1038/nchembio.542
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DOI: https://doi.org/10.1038/nchembio.542