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Related Experiment Video

Updated: Jun 1, 2026

Evaluation of LC3-II Release via Extracellular Vesicles in Relation to the Accumulation of Intracellular LC3-positive Vesicles

Evaluation of LC3-II Release via Extracellular Vesicles in Relation to the Accumulation of Intracellular LC3-positive Vesicles

Published on: October 18, 2024

ErbB2-associated changes in the lysosomal proteome.

Jesper Nylandsted¹, Andrea C Becker, Jakob Bunkenborg

¹Apoptosis Department and Centre for Genotoxic Stress Research, Institute of Cancer Biology, Danish Cancer Society, Strandboulevarden 49, Copenhagen, Denmark.

|June 16, 2011

Summary

This summary is machine-generated.

This study presents a new antibody-based method for purifying intact lysosomes, crucial for cellular recycling. This technique enables better analysis of lysosomes in diseases like cancer, unlike older methods.

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Area of Science:

Cell Biology
Molecular Biology
Proteomics

Background:

Lysosomes are vital organelles for cellular waste removal and macromolecule turnover.
Current lysosome purification methods (density gradient centrifugation, iron-loaded organelle magnetic purification) are limited by changes in lysosome density and stability in disease states.
These limitations hinder the comparative analysis of normal versus pathological lysosomes.

Purpose of the Study:

To develop an efficient and robust method for purifying intact lysosomes suitable for comparative analysis.
To characterize the lysosomal membrane proteome using quantitative mass spectrometry.
To investigate alterations in the lysosomal proteome associated with cancer, specifically ErbB2 oncogene expression.

Main Methods:

Development of a magnetic immunoprecipitation technique using antibodies against the vacuolar-type H(+) -ATPase for lysosome purification.
Quantitative mass spectrometry (MS)-based proteomics for identifying proteins in purified lysosomal membranes.
Comparative proteomic analysis of lysosomes from cells with and without ectopic ErbB2 oncogene expression.

Main Results:

Successful purification of intact lysosomes using antibody-based magnetic immunoprecipitation.
Identification of 60 proteins in the lysosomal membrane proteome, with most previously linked to lysosomes.
Significant alterations in the lysosomal membrane proteome were observed upon ectopic ErbB2 expression, including increased CD63, S100A11, and ferritin heavy chain levels.

Conclusions:

Antibody-based purification of lysosomes is an effective method for characterizing lysosomes, particularly in pathological conditions.
This method overcomes limitations of traditional techniques, enabling better study of lysosomes in diseases like cancer.
The study reveals specific protein changes in the lysosomal membrane associated with ErbB2-driven metastasis.