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Cross-link assisted spatial proteomics to map sub-organelle proteomes and membrane protein topology

ABSTRACT: The specific functions of cellular organelles and sub-compartments depend on their protein content, which can be characterized by spatial proteomics approaches. However, many spatial proteomics methods are limited in their ability to resolve organellar sub-compartments, profile multiple sub-compartments in parallel, and/or characterize membrane-associated proteomes. Here, we develop a cross-linking assisted spatial proteomics (CLASP) strategy that addresses these shortcomings. Using human mitochondria as a model system, we show that CLASP can elucidate spatial proteomes of all mitochondrial sub-compartments and provide topological insight into the mitochondrial membrane proteome in a single experiment. Biochemical and imaging-based follow-up studies demonstrate that CLASP allows discovering mitochondria-associated proteins and revising previous protein sub-compartment localization and membrane topology data. This study extends the scope of cross-linking mass spectrometry beyond protein structure and interaction analysis towards spatial proteomics, establishes a method for concomitant profiling of sub-organelle and membrane proteomes, and provides a resource for mitochondrial spatial biology.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Homo Sapiens (human) Mus Musculus (mouse)

TISSUE(S): Brain, Cell Culture

SUBMITTER: Zhu Ying

LAB HEAD: Fan Liu

PROVIDER: PXD032132 | Pride | 2024-02-21

REPOSITORIES: Pride

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			Action	DRS
	F1_20190509_YZ_04_1.raw	Raw
	F1_20190509_YZ_04_2.raw	Raw
	F1_20190509_YZ_04_3.raw	Raw
	F1_20190517_YZ_05_1.raw	Raw
	F1_20190517_YZ_05_2.raw	Raw

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