The proteome of Nicotiana benthamiana is shaped by extensive protein processing

© 2024 The Authors. New Phytologist © 2024 New Phytologist Foundation.

Détails bibliographiques
Publié dans:The New phytologist. - 1979. - 243(2024), 3 vom: 10. Aug., Seite 1034-1049
Auteur principal: Zheng, Kaijie (Auteur)
Autres auteurs: Lyu, Joy C, Thomas, Emma L, Schuster, Mariana, Sanguankiattichai, Nattapong, Ninck, Sabrina, Kaschani, Farnusch, Kaiser, Markus, van der Hoorn, Renier A L
Format: Article en ligne
Langue:English
Publié: 2024
Accès à la collection:The New phytologist
Sujets:Journal Article Nicotiana benthamiana agroinfiltration molecular weight protein migration protein processing Proteome Plant Proteins Carboxylic Ester Hydrolases EC 3.1.1.- plus... pectinesterase EC 3.1.1.11 Lipase EC 3.1.1.3 Peptide Hydrolases EC 3.4.- Glycoside Hydrolases EC 3.2.1.-
Description
Résumé:© 2024 The Authors. New Phytologist © 2024 New Phytologist Foundation.
Processing by proteases irreversibly regulates the fate of plant proteins and hampers the production of recombinant proteins in plants, yet only few processing events have been described in agroinfiltrated Nicotiana benthamiana, which has emerged as the main transient protein expression platform in plant science and molecular pharming. Here, we used in-gel digests and mass spectrometry to monitor the migration and topography of 5040 plant proteins within a protein gel. By plotting the peptides over the gel slices, we generated peptographs that reveal where which part of each protein was detected within the protein gel. These data uncovered that 60% of the detected proteins have proteoforms that migrate at lower than predicted molecular weights, implicating extensive proteolytic processing. This analysis confirms the proteolytic removal and degradation of autoinhibitory prodomains of most but not all proteases, and revealed differential processing within pectinemethylesterase and lipase families. This analysis also uncovered intricate processing of glycosidases and uncovered that ectodomain shedding might be common for a diverse range of receptor-like kinases. Transient expression of double-tagged candidate proteins confirmed processing events in vivo. This large proteomic dataset implicates an elaborate proteolytic machinery shaping the proteome of N. benthamiana
Description:Date Completed 04.07.2024
Date Revised 24.10.2024
published: Print-Electronic
Citation Status MEDLINE
ISSN:1469-8137
DOI:10.1111/nph.19891