Górna / Klimecka / Izert-Nowakowska Degrons

Methods and Protocols
Erscheinungsjahr 2026
ISBN: 978-1-0716-5508-5
Verlag: Springer US
Format: PDF
Kopierschutz: 1 - PDF Watermark

E-Book, Englisch, 260 Seiten

Reihe: Springer Protocols

This detailed volume provides protocols for the identification, characterization, and application of degrons, central to controlling protein abundance and cellular homeostasis, in a range of organisms from bacteria to humans. Beginning with reporter assays and tools used in human cells to study the involvement of the ubiquitin-proteasome system or ubiquitin-independent degradation pathways, the book continues by exploring methods for chemical and enzymatic synthesis, characterization of degrons and their mimics, methods for other selected eukaryotes, including plants and employing inducible degradation in nematodes and trypanosomes, as well as methods used for studying N- and C-degrons in bacteria. Written for the highly successful series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step and readily reproducible laboratory protocols, and tips for troubleshooting and avoiding known pitfalls.

Authoritative and practical, serves as an ideal guide for researchers working on protein degradation or using degrons to study or control protein function.

Zielgruppe

Professional/practitioner

Autoren/Hrsg.

Górna, Maria W.

Klimecka, Maria M.

Izert-Nowakowska, Matylda A.

Inhaltsverzeichnis

HiBiT-RR and nLucK0 Tagging Systems for Monitoring Targeted Protein Degradation.- Identification of a Ubiquitin-Independent Degron by a Reporter Assay.- Utilization of Functional Degradation Pathways for the Characterization of Ubiquitin-Proteasome System Tool Compounds.- UbiSite Approach: Global Site-Specific Profiling of Canonical and Non-Canonical Ubiquitination in Cells and Tissues.- Chemical and Enzymatic Approaches to C-Terminal Cyclic Imide Formation.- Design and Semisynthesis of Ubiquitin Extension Probes.- Biophysical and Cellular Assays for the Evaluation of Peptidomimetic Inhibitors Based on Degron Sequences.- Multicolor Live-Cell mRNA Imaging Using RNA-Regulated Destabilization Domains.- Dynamic Control of Cell Signaling with Optogenetic Modulation of Protein Abundance in Living Mammalian Cells.- Reporter Gene Assay for Monitoring BCL6 Reactivation.- Studying Glycan-Dependent ERAD of Misfolded Glycoproteins in Plants.- Protein Depletion in C. elegans Using the Auxin-Inducible Degradation System.- An Auxin-Inducible Degron System for Trypanosomes.- Degrade Green Fluorescent Protein (deGradFP) Method in Trypanosoma brucei.- Production of Purified Proteins Tagged with ssrA-Derived Degrons and Their Degradation by the Escherichia coli ClpXP System.- Evaluation of Degron Motifs in Escherichia coli Using a Fluorescent Reporter.- Assessment of Bacterial N-Degron Proteolysis by a Dual-Fluorescence Reporter.- SsrA-Based Targeted Protein Degradation in Escherichia coli.