A bifunctional asparaginyl endopeptidase efficiently catalyzes both cleavage and cyclization of cyclic trypsin inhibitors
Autor: | Quentin Kaas, Edward K. Gilding, David J. Craik, Fong Yang Looi, Lai Yue Chan, Thomas Durek, Kuok Yap, Junqiao Du, Aaron G. Poth, Fabian B. H. Rehm |
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Jazyk: | angličtina |
Rok vydání: | 2020 |
Předmět: |
0301 basic medicine
Models Molecular Stereochemistry Trypsin inhibitor Science General Physics and Astronomy 010402 general chemistry Protein Engineering 01 natural sciences Peptides Cyclic General Biochemistry Genetics and Molecular Biology Article Substrate Specificity 03 medical and health sciences Cyclotides medicine Enzyme kinetics Amino Acid Sequence lcsh:Science Plant Proteins chemistry.chemical_classification Multidisciplinary General Chemistry Protein engineering Proteases Hydrogen-Ion Concentration Trypsin Cyclic peptide Endopeptidase Recombinant Proteins 0104 chemical sciences Cyclotide Enzymes Cysteine Endopeptidases Kinetics 030104 developmental biology chemistry Cyclization Biocatalysis lcsh:Q Peptides Trypsin Inhibitors medicine.drug |
Zdroj: | Nature Communications, Vol 11, Iss 1, Pp 1-11 (2020) Nature Communications |
ISSN: | 2041-1723 |
Popis: | Asparaginyl endopeptidases (AEPs) catalyze the key backbone cyclization step during the biosynthesis of plant-derived cyclic peptides. Here, we report the identification of two AEPs from Momordica cochinchinensis and biochemically characterize MCoAEP2 that catalyzes the maturation of trypsin inhibitor cyclotides. Recombinantly produced MCoAEP2 catalyzes the backbone cyclization of a linear cyclotide precursor (MCoTI-II-NAL) with a kcat/Km of 620 mM−1 s−1, making it one of the fastest cyclases reported to date. We show that MCoAEP2 can mediate both the N-terminal excision and C-terminal cyclization of cyclotide precursors in vitro. The rate of cyclization/hydrolysis is primarily influenced by varying pH, which could potentially control the succession of AEP-mediated processing events in vivo. Furthermore, MCoAEP2 efficiently catalyzes the backbone cyclization of an engineered MCoTI-II analog with anti-angiogenic activity. MCoAEP2 provides enhanced synthetic access to structures previously inaccessible by direct chemistry approaches and enables the wider application of trypsin inhibitor cyclotides in biotechnology applications. Asparaginyl endopeptidases (AEPs) catalyze the cyclization step during the biosynthesis of cyclic peptides in plants. Here, the authors report a recombinantly produced AEP that catalyzes the backbone cyclization of a linear cyclotide precursor and an engineered analog with high efficiency and in a pH-dependent manner. |
Databáze: | OpenAIRE |
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