Proteomics

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A scalable multiplexed platform to accelerate protein interactome discovery

ABSTRACT: Chromatographic fractionation-mass spectrometry (CF-MS) allows for mapping native protein-protein interactions and macromolecular complexes on a proteome-scale but is tedious and resource intensive for comparative analysis. Here, we present a more parsimonious and reproducible multiplexing CF-MS platform for measuring multi-protein assemblies across different experimental samples simultaneously with an order-of-magnitude faster than previous approaches. We used this innovative strategy to elucidate protein networks in control and breast cancer cells models in parallel within two weeks of total MS instrument time.

INSTRUMENT(S): Q Exactive HF

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Epithelial Cell, Cell Culture

DISEASE(S): Breast Cancer

SUBMITTER: Pierre Havugimana  

LAB HEAD: Andrew Emili

PROVIDER: PXD027704 | Pride | 2022-08-03

REPOSITORIES: Pride

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Scalable multiplex co-fractionation/mass spectrometry platform for accelerated protein interactome discovery.

Havugimana Pierre C PC   Goel Raghuveera Kumar RK   Phanse Sadhna S   Youssef Ahmed A   Padhorny Dzmitry D   Kotelnikov Sergei S   Kozakov Dima D   Emili Andrew A  

Nature communications 20220713 1

Co-fractionation/mass spectrometry (CF/MS) enables the mapping of endogenous macromolecular networks on a proteome scale, but current methods are experimentally laborious, resource intensive and afford lesser quantitative accuracy. Here, we present a technically efficient, cost-effective and reproducible multiplex CF/MS (mCF/MS) platform for measuring and comparing, simultaneously, multi-protein assemblies across different experimental samples at a rate that is up to an order of magnitude faster  ...[more]

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