Dataset Information

On-chip AP-MS: Interaction proteomics on a microfluidics platform

ABSTRACT: Essentially all cellular processes are orchestrated by protein-protein interactions (PPIs). In recent years, affinity purification coupled to mass spectrometry (AP-MS) has been the preferred method to identify cellular PPIs. Here we present a microfluidics-based (AP-MS) workflow to identify PPIs using minute amounts of input material. The use of this automated platform allowed us to identify the human cohesin, CCC and Mediator complex from as little as 4 micrograms of input lysate, representing a 50─100-fold downscaling compared to regular microcentrifuge tube-based protocols. We show that our platform can be used to affinity purify tagged baits as well as native cellular proteins and their interaction partners. As such, our method holds great promise for future biological and clinical AP-MS applications in which sample amounts are limited.

INSTRUMENT(S): Orbitrap Fusion ETD

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Cristina Furlan

LAB HEAD: Michiel Vermeulen

PROVIDER: PXD012800 | Pride | 2019-04-02

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	1.5M_300K_60K_RESULTS.rar	Other
	1.5m_300k_60k_SMC1A_PD_4ug.rar	Other
	12K_Ab6556_SMC1A_RESULTS.rar	Other
	12kSC_BUBR1_mit_PD.rar	Other
	12kSC_BUBR1_mit_PD_RESULTS.rar	Other

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Publications

Miniaturised interaction proteomics on a microfluidic platform with ultra-low input requirements.

Furlan Cristina C Dirks René A M RAM Thomas Peter C PC Jones Robert C RC Wang Jing J Lynch Mark M Marks Hendrik H Vermeulen Michiel M

Nature communications 20190404 1

Essentially all cellular processes are orchestrated by protein-protein interactions (PPIs). In recent years, affinity purification coupled to mass spectrometry (AP-MS) has been the preferred method to identify cellular PPIs. Here we present a microfluidic-based AP-MS workflow, called on-chip AP-MS, to identify PPIs using minute amounts of input material. By using this automated platform we purify the human Cohesin, CCC and Mediator complexes from as little as 4 micrograms of input lysate, repres ...[more]

PMID: 30948724

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