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Lysosomal proteomics by affinity purification-mass spectrometry

ABSTRACT: The aims of this project is to establish a comprehensive proteomic map of the lysosomes using isolated lysosomes via affinity purification. Affinity-purified mitochondria was used as a control to demonstrate the organelle specificity. In this study, we identified several lipid transporters that are associated with the lysosome and further characterized their roles in sterol-dependent mTORC1 regulation.

INSTRUMENT(S): LTQ Orbitrap Elite, Q Exactive

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Epithelial Cell

SUBMITTER: Chun-Yan Lim

LAB HEAD: Roberto Zoncu

PROVIDER: PXD014733 | Pride | 2019-09-27

REPOSITORIES: Pride

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ER-lysosome contacts enable cholesterol sensing by mTORC1 and drive aberrant growth signalling in Niemann-Pick type C.

Lim Chun-Yan CY Davis Oliver B OB Shin Hijai R HR Zhang Justin J Berdan Charles A CA Jiang Xuntian X Counihan Jessica L JL Ory Daniel S DS Nomura Daniel K DK Zoncu Roberto R

Nature cell biology 20190923 10

Cholesterol activates the master growth regulator, mTORC1 kinase, by promoting its recruitment to the surface of lysosomes by the Rag guanosine triphosphatases (GTPases). The mechanisms that regulate lysosomal cholesterol content to enable mTORC1 signalling are unknown. Here, we show that oxysterol binding protein (OSBP) and its anchors at the endoplasmic reticulum (ER), VAPA and VAPB, deliver cholesterol across ER-lysosome contacts to activate mTORC1. In cells lacking OSBP, but not other VAP-in ...[more]

PMID: 31548609

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