The macrocyclizing protease butelase 1 remains autocatalytic and reveals the structural basis for ligase activity

The Plant Journal : for Cell and Molecular Biology
Amy M JamesJoshua S Mylne

Abstract

Plant asparaginyl endopeptidases (AEPs) are expressed as inactive zymogens that perform maturation of seed storage protein upon cleavage-dependent autoactivation in the low-pH environment of storage vacuoles. The AEPs have attracted attention for their macrocyclization reactions, and have been classified as cleavage or ligation specialists. However, we have recently shown that the ability of AEPs to produce either cyclic or acyclic products can be altered by mutations to the active site region, and that several AEPs are capable of macrocyclization given favorable pH conditions. One AEP extracted from Clitoria ternatea seeds (butelase 1) is classified as a ligase rather than a protease, presenting an opportunity to test for loss of cleavage activity. Here, making recombinant butelase 1 and rescuing an Arabidopsis thaliana mutant lacking AEP, we show that butelase 1 retains cleavage functions in vitro and in vivo. The in vivo rescue was incomplete, consistent with some trade-off for butelase 1 specialization toward macrocyclization. Its crystal structure showed an active site with only subtle differences from cleaving AEPs, suggesting the many differences in its peptide-binding region are the source of its efficient macrocyclizat...Continue Reading

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Citations

Apr 21, 2019·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Shangwen Luo, Shi-Hui Dong
Nov 5, 2019·The New Phytologist·Kenji YamadaIkuko Hara-Nishimura
Jul 28, 2019·Scientific Reports·Karen S HarrisMarilyn A Anderson
Jun 22, 2019·Journal of Experimental Botany·Simon StaelMoritz K Nowack
Sep 2, 2020·The Journal of Biological Chemistry·Ross W ChelohaHidde L Ploegh
Nov 28, 2019·Chemical Reviews·Robert E Thompson, Tom W Muir
Feb 19, 2021·Medicinal Research Reviews·Jingjing ZhangChengchao Xu
Feb 27, 2021·Journal of the American Chemical Society·Elise A NaudinVladimir Torbeev
Mar 6, 2021·Biochemical Society Transactions·Samuel G NonisJoshua S Mylne
Mar 10, 2021·Chembiochem : a European Journal of Chemical Biology·Fabian B H RehmDavid J Craik
Jun 8, 2021·Journal of the American Chemical Society·Dingpeng ZhangChuan-Fa Liu
Oct 2, 2019·Journal of the American Chemical Society·Fabian B H RehmHidde L Ploegh

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