s41467-018-04669-9.pdf (1.78 MB)
Download fileMolecular basis for the production of cyclic peptides by plant asparaginyl endopeptidases
journal contribution
posted on 2021-02-11, 23:49 authored by MA Jackson, EK Gilding, Thomas ShafeeThomas Shafee, Karen HarrisKaren Harris, Q Kaas, Simon PoonSimon Poon, K Yap, H Jia, Rosemary GuarinoRosemary Guarino, LY Chan, T Durek, Marilyn AndersonMarilyn Anderson, DJ Craik© 2018 The Author(s). Asparaginyl endopeptidases (AEPs) are proteases that have crucial roles in plant defense and seed storage protein maturation. Select plant AEPs, however, do not function as proteases but as transpeptidases (ligases) catalyzing the intra-molecular ligation of peptide termini, which leads to peptide cyclization. These ligase-type AEPs have potential biotechnological applications ranging from in vitro peptide engineering to plant molecular farming, but the structural features enabling these enzymes to catalyze peptide ligation/cyclization rather than proteolysis are currently unknown. Here, we compare the sequences, structures, and functions of diverse plant AEPs by combining molecular modeling, sequence space analysis, and functional testing in planta. We find that changes within the substrate-binding pocket and an adjacent loop, here named the "marker of ligase activity", together play a key role for AEP ligase efficiency. Identification of these structural determinants may facilitate the discovery of more ligase-type AEPs and the engineering of AEPs with tailored catalytic properties.
Funding
The pEAQ vectors were kindly provided by Prof. George Lomonossoff at the John Innes Centre and Plant Bioscience Ltd. We acknowledge funding from the Australian Research Council (ARC Laureate Fellowship (FL150100146) to D.J.C. and ARC grant DP150100443 to D.J.C., E.K.G., and T.D.). This research was also supported by the 2015 Ramaciotti Biomedical Research Award to D.J.C. and M.A.A. and by Hexima Ltd.
Australian Research Council (ARC) | FL150100146
Australian Research Council (ARC) | DP150100443
Ramaciotti Biomedical Research Award
Hexima Ltd.
History
Publication Date
2018-06-20Journal
Nature CommunicationsVolume
9Issue
1Article Number
2411Pagination
12p. (p. 1-12)Publisher
Springer NatureISSN
2041-1723Rights Statement
The Author reserves all moral rights over the deposited text and must be credited if any re-use occurs. Documents deposited in OPAL are the Open Access versions of outputs published elsewhere. Changes resulting from the publishing process may therefore not be reflected in this document. The final published version may be obtained via the publisher’s DOI. Please note that additional copyright and access restrictions may apply to the published version.Publisher DOI
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Science & TechnologyMultidisciplinary SciencesScience & Technology - Other TopicsKNOTTED PROTEINSHOST-DEFENSECYSTINE KNOTCELL-DEATHKALATA B1CYCLOTIDESBIOSYNTHESISLEGUMAINFAMILYCYCLIZATIONPlantsPlants, Genetically ModifiedPeptides, CyclicCysteine EndopeptidasesPlant ProteinsRNA, PlantSequence Analysis, RNAModels, Molecular