Substrate-binding glycine residues are major determinants for hydrolase and ligase activity of plant legumains

Substrate-binding glycine residues are major determinants for hydrolase and ligase activity of plant legumains

© 2023 The Authors. New Phytologist © 2023 New Phytologist Foundation.

Bibliographische Detailangaben
Veröffentlicht in:The New phytologist. - 1979. - 238(2023), 4 vom: 01. Mai, Seite 1534-1545
1. Verfasser: Hemu, Xinya (VerfasserIn)
Weitere Verfasser: Chan, Ning-Yu, Liew, Heng Tai, Hu, Side, Zhang, Xiaohong, Serra, Aida, Lescar, Julien, Liu, Chuan-Fa, Tam, James P
Format: Online-Aufsatz
Sprache:English
Veröffentlicht: 2023
Zugriff auf das übergeordnete Werk:The New phytologist
Schlagworte:Journal Article Research Support, Non-U.S. Gov't asparaginyl endopeptidase butelase ligase-activity determinant peptide asparaginyl ligase plant legumain asparaginylendopeptidase EC 3.4.22.34 Plant Proteins mehr... Glycine TE7660XO1C Cysteine Endopeptidases EC 3.4.22.- Ligases EC 6.-
Beschreibung
Zusammenfassung:© 2023 The Authors. New Phytologist © 2023 New Phytologist Foundation.
Peptide asparaginyl ligases (PALs) are useful tools for precision modifications of proteins and live-cell surfaces by ligating peptides after Asn/Asp (Asx). They share high sequence and structural similarity to plant legumains that are generally known as asparaginyl endopeptidases (AEPs), thus making it challenging to identify PALs from AEPs. In this study, we investigate 875 plant species from algae to seed plants with available sequence data in public databases to identify new PALs. We conducted evolutionary trace analysis on 1500 plant legumains, including eight known PALs, to identify key residues that could differentiate ligases and proteases, followed by recombinant expression and functional validation of 16 novel legumains. Previously, we showed that the substrate-binding sequences flanking the catalytic site can strongly influence the enzymatic direction of a legumain and which we named as ligase-activity determinants (LADs). Here, we show that two conserved substrate-binding Gly residues of LADs are critical, but negative determinants for ligase activity. Our results suggest that specific glycine residues are molecular determinants to identify PALs and AEPs as two different legumain subfamilies, accounting for c. 1% and 88%, respectively
Beschreibung:Date Completed 14.04.2023
Date Revised 21.04.2023
published: Print-Electronic
Citation Status MEDLINE
ISSN:1469-8137
DOI:10.1111/nph.18841