EMD-51701

Tomography
EMD-51701 Deposition: 02/10/2024
Map released: 05/02/2025
Last modified: 05/02/2025
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EMD-51701

In situ cryo-electron tomogram of a multi-lamellar vesicle in a NPC2-/- HeLa cell. #1

EMD-51701

Tomography
EMD-51701 Deposition: 02/10/2024
Map released: 05/02/2025
Last modified: 05/02/2025
Overview Sample Experiment Validation Volume Browser Additional data Links
Sample Organism: Homo sapiens
Sample: HeLa TMEM192-3xHA NPC2-/-

Deposition Authors: Kraus F , He Y , Swarup S , Overmyer KA , Jiang Y , Brenner J , Capitanio C , Bieber A , Jen A , Nightingale NM , Anderson BJ , Lee C , Paulo JA , Smith IR , Plitzko JM , Gygi SP , Schulman BA , Wilfling F , Coon JJ , Harper JW
Global cellular proteo-lipidomic profiling of diverse lysosomal storage disease mutants using nMOST.
PUBMED: 39841834
DOI: doi:10.1126/sciadv.adu5787
ISSN: 2375-2548
Abstract:
Lysosomal storage diseases (LSDs) comprise ~50 monogenic disorders marked by the buildup of cellular material in lysosomes, yet systematic global molecular phenotyping of proteins and lipids is lacking. We present a nanoflow-based multiomic single-shot technology (nMOST) workflow that quantifies HeLa cell proteomes and lipidomes from over two dozen LSD mutants. Global cross-correlation analysis between lipids and proteins identified autophagy defects, notably the accumulation of ferritinophagy substrates and receptors, especially in NPC1-/- and NPC2-/- mutants, where lysosomes accumulate cholesterol. Autophagic and endocytic cargo delivery failures correlated with elevated lysophosphatidylcholine species and multilamellar structures visualized by cryo-electron tomography. Loss of mitochondrial cristae, MICOS complex components, and OXPHOS components rich in iron-sulfur cluster proteins in NPC2-/- cells was largely alleviated when iron was provided through the transferrin system. This study reveals how lysosomal dysfunction affects mitochondrial homeostasis and underscores nMOST as a valuable discovery tool for identifying molecular phenotypes across LSDs.