EMD-51701
In situ cryo-electron tomogram of a multi-lamellar vesicle in a NPC2-/- HeLa cell. #1
EMD-51701
Tomography
Map released: 05/02/2025
Last modified: 05/02/2025
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
Sample: HeLa TMEM192-3xHA NPC2-/-
Deposition Authors: Kraus F
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Global cellular proteo-lipidomic profiling of diverse lysosomal storage disease mutants using nMOST.
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
(2025) Sci Adv , 11 , eadu5787 - eadu5787
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(2025) Sci Adv , 11 , eadu5787 - eadu5787
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.
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.