MemPrep, a New Technology for Isolating Organellar Membranes Provides Fingerprints of Lipid Bilayer Stress

Overview

Journal EMBO J

Specialties Biotechnology
Molecular Biology

Date 2024 Mar 16

PMID 38491296

Authors

John Reinhard

Leonhard Starke

Christian Klose

Per Haberkant

Henrik Hammaren

Frank Stein

Ofir Klein

Charlotte Berhorst

Heike Stumpf

James P Saenz

Jochen Hub

Maya Schuldiner

Robert Ernst

Affiliations

Soon will be listed here.

Abstract

Biological membranes have a stunning ability to adapt their composition in response to physiological stress and metabolic challenges. Little is known how such perturbations affect individual organelles in eukaryotic cells. Pioneering work has provided insights into the subcellular distribution of lipids in the yeast Saccharomyces cerevisiae, but the composition of the endoplasmic reticulum (ER) membrane, which also crucially regulates lipid metabolism and the unfolded protein response, remains insufficiently characterized. Here, we describe a method for purifying organelle membranes from yeast, MemPrep. We demonstrate the purity of our ER membrane preparations by proteomics, and document the general utility of MemPrep by isolating vacuolar membranes. Quantitative lipidomics establishes the lipid composition of the ER and the vacuolar membrane. Our findings provide a baseline for studying membrane protein biogenesis and have important implications for understanding the role of lipids in regulating the unfolded protein response (UPR). The combined preparative and analytical MemPrep approach uncovers dynamic remodeling of ER membranes in stressed cells and establishes distinct molecular fingerprints of lipid bilayer stress.

Citing Articles

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