Projects per year
Abstract
Over 30 years ago, an intriguing post-translational modification was found responsible for creating concanavalin A (conA), a carbohydrate-binding protein from jack bean (Canavalia ensiformis) seeds and a common carbohydrate chromatography reagent. ConA biosynthesis involves what was then an unprecedented rearrangement in amino-acid sequence, whereby the N-terminal half of the gene-encoded conA precursor is swapped to become the C-terminal half of conA. Asparaginyl endopeptidase (AEP) was shown to be involved, but its mechanism was not fully elucidated. To understand the structural basis and consequences of circular permutation, we generated recombinant jack bean conA precursor (pro-conA) plus jack bean AEP (CeAEP1) and solved crystal structures for each to 2.1 Å and 2.7 Å, respectively. By reconstituting conA biosynthesis in vitro, we prove CeAEP1 alone can perform both cleavage and cleavage-coupled transpeptidation to form conA. CeAEP1 structural analysis reveals how it is capable of carrying out both reactions. Biophysical assays illustrated that pro-conA is less stable than conA. This observation was explained by fewer intermolecular interactions between subunits in the pro-conA crystal structure and consistent with a difference in the prevalence for tetramerisation in solution. These findings elucidate the consequences of circular permutation in the only post-translation example known to occur in nature.
Original language | English |
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Pages (from-to) | 2794 - 2811 |
Number of pages | 18 |
Journal | The Plant Cell |
Volume | 33 |
Issue number | 8 |
DOIs | |
Publication status | Published - Aug 2021 |
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Dive into the research topics of 'Structural and biochemical analyses of concanavalin A circular permutation by jack bean asparaginyl endopeptidase'. Together they form a unique fingerprint.Projects
- 2 Finished
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Structure-based investigations into new modes of action for herbicides
Haywood, J.
ARC Australian Research Council
2/04/18 → 1/05/21
Project: Research
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How scissors learn to glue: the catalysis of ligation by proteases
Mylne, J. & Tawfik, D.
ARC Australian Research Council
1/01/16 → 31/12/18
Project: Research