Plant Proteostasis: Methods and Protocols

by L. Maria Lois (Editor), Marco Trujillo (Editor)

Overview

This second edition compiles state-of-art prools to study the molecular mechanisms that determine proteome composition and contribute to maintaining protein homeostasis. Chapters describe methods that range from the in vitro analysis of enzyme cascades to real-time in vivo studies. Written in the successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible prools, and notes on troubleshooting and avoiding known pitfalls.

Authoritative and cutting-edge, Plant Proteostasis: Methods and Prools, Second Edition aims to be a useful practical guide to researches to help further their study in this field.

Product Details

ISBN-13:	9781071627860
Publisher:	Springer US
Publication date:	11/23/2023
Series:	Methods in Molecular Biology , #2581
Edition description:	2nd ed. 2023
Pages:	415
Product dimensions:	7.01(w) x 10.00(h) x (d)

Table of Contents

Approaches to Determine Protein Ubiquitination Residue Types.- Immunoprecipitation of Cullin Ring Ligases (CRLs) in Arabidopsis Thaliana Seedlings.- Radioligand Binding Assays for Determining Dissociation Constants of Phytohormone Receptors.- Measuring the Enzyme Activity of Arabidopsis Deubiquitylating Enzymes.- Fluorescent Reporters for Ubiquitin-Dependent Proteolysis in Plants.- Generation of Artificial N-end Rule Substrate Proteins In Vivo and In Vitro.- Peptide Arrays for Binding Studies of E3 Ubiquitin Ligases.- SUMO Chain Formation by Plants Enzymes.- Kinetic Analysis of Plant SUMO Conjugation Machinery.- Expression, Purification, and Enzymatic Analysis Of Plant SUMO Proteases.- Quantitative Analysis of Subcellular Distribution of the SUMO Conjugation System by Confocal Microscopy Imaging.- Biochemical Analysis of Autophagy in Algae and Plants by Monitoring the Electrophoretic Mobility of ATG8.- Detection of Autophagy in Plants by Fluorescence Microscopy.- Prools for Studying Protein Stability in a Arabidopsis Protoplast Transient Expression System.- Detection and Quantification of Protein Aggregates in Plants.- Determination of Protein Carbonylation and Proteasome Activity in Seeds.- Isobaric Tag for Relative and Absolute Quantitation (iTRAQ)-based Protein Profiling in Plants.-Use of a Phosphatidylinositol phosphate Affinity Chromatography (PIP - chromatography) for the Isolation of Proteins Involved in Protein Quality Control and Proteostasis Mechanisms in Plants.- In vivo Radiolabeling of Arabidopsis Leaves and Separation of Thylakoid Membrane Complexes by Blue Native PAGE.- Normalized Quantitative Western Blotting Based on Standardized Fluorescent Labeling.- Sequence Search and Comparative Genomic Analysis of SUMO-Activating Enzymes using CoGe.- Studying Evolutionary Dynamics of Gene Families Encoding SUMO-Activating Enzymes with SeaView and ProtTest.- Bioinformatics Tools for Exploring the SUMO Gene Network.