Visualization of Lysosomal Membrane Proteins by Cryo Electron Tomography

Authors

Bridget M McVeigh
José J De Jesús-Pérez
Dirk H Siepe
Prerana Gogoi
Shrawan Kumar Mageswaran
Marian Kalocsay
Elaine M Mihelc
Vera Y Moiseenkova-Bell

Language

English

Publication Date

10-17-2025

Journal

Nature Communications

DOI

10.1038/s41467-025-64314-0

PMID

41107240

PMCID

PMC12534608

PubMedCentral® Posted Date

10-17-2025

PubMedCentral® Full Text Version

Post-print

Abstract

Lysosomes are essential organelles for cellular homeostasis and signaling, with dysfunction linked to neurological disorders, lysosomal storage diseases, and cancer. While proteomics has advanced our understanding of lysosomal composition, the structural characterization of lysosomal membrane proteins in their native environment remains a significant challenge. Here, we developed a cryo electron tomography workflow to visualize lysosomal membrane proteins within intact, native lysosomal membranes. We isolated endolysosomes by independently targeting two lysosomal membrane proteins, transient receptor potential mucolipin 1 and transmembrane protein 192, enriching organelles that exhibited the expected morphology and proteomic composition of the endolysosomal system. Sub-tomogram averaging enabled the structural refinement of key membrane and membrane-associated proteins, including V-ATPase, Flotillin, and Clathrin, directly within the lysosomal membrane, revealing their heterogeneous distribution across endolysosomal organelles. By integrating proteomics with structural biology, our workflow establishes a powerful platform for studying lysosomal membrane protein function in health and disease, paving the way for future discoveries in membrane-associated lysosomal mechanisms.

Keywords

Lysosomes, Humans, Cryoelectron Microscopy, Electron Microscope Tomography, Membrane Proteins, Lysosomal Membrane Proteins, Proteomics, Transient Receptor Potential Channels, Intracellular Membranes, Vacuolar Proton-Translocating ATPases, Clathrin, Animals, Cryoelectron tomography, Lysosomes, Cell biology

Published Open-Access

yes

Recommended Citation

McVeigh, Bridget M; De Jesús-Pérez, José J; Siepe, Dirk H; et al., "Visualization of Lysosomal Membrane Proteins by Cryo Electron Tomography" (2025). Faculty, Staff and Student Publications. 5946.
https://digitalcommons.library.tmc.edu/uthgsbs_docs/5946