Targeted Protein Unfolding at the Golgi Apparatus

Jasmin Schillinger; Doris Hellerschmied

doi:10.1007/978-1-0716-2639-9_39

Targeted Protein Unfolding at the Golgi Apparatus

Methods Mol Biol. 2023:2557:645-659. doi: 10.1007/978-1-0716-2639-9_39.

Authors

Jasmin Schillinger¹, Doris Hellerschmied²

Affiliations

¹ Faculty of Biology, Center of Medical Biotechnology, University Duisburg-Essen, Essen, Germany.
² Faculty of Biology, Center of Medical Biotechnology, University Duisburg-Essen, Essen, Germany. doris.hellerschmied@uni-due.de.

PMID: 36512243
DOI: 10.1007/978-1-0716-2639-9_39

Abstract

Maintaining protein homeostasis (proteostasis) is vital to cellular and organismal health. How the Golgi apparatus, the central protein maturation and sorting station in the cell, manages misfolded proteins to maintain proteostasis is still poorly understood. Here we present a strategy for targeted protein unfolding at the Golgi that enables studying Golgi-related protein quality control and stress-signaling pathways. Targeted protein unfolding is induced by small molecule-based chemical biology approaches-hydrophobic tagging and the use of a destabilization domain. Imaging studies allow visualizing quality control (QC) phenotypes, such as the formation of QC carriers and Golgi-to-endoplasmic reticulum trafficking, and correlating these phenotypes with other trafficking processes.

Keywords: Chemical biology tools; Destabilization domain; HaloTag fusion proteins; Hydrophobic tagging; SNAP-TMR pulse labeling; Targeted protein unfolding; Trimethoprim.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Endoplasmic Reticulum* / metabolism
Golgi Apparatus* / metabolism
Protein Transport
Protein Unfolding