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Rapid profiling of protein complex re-organization in perturbed systems

ABSTRACT: Protein complexes constitute the primary functional modules of cellular activity. To respond to perturbations, complexes undergo changes in their abundance, subunit composition or state of modification. Understanding the function of biological systems requires global strategies to capture this contextual state information on protein complexes and interaction networks. Methods based on co-fractionation paired with mass spectrometry have demonstrated the capability for deep biological insight but the scope of studies using this approach has been limited by the large measurement time per biological sample and challenges with data analysis. As such, there has been little uptake of this strategy beyond a few expert labs into the broader life science community despite rich biological information content. We present a rapid integrated experimental and computational workflow to assess the re-organization of protein complexes across multiple cellular states. It enables complex experimental designs requiring increased sample/condition numbers. The workflow combines short gradient chromatography and DIA/SWATH mass spectrometry with a data analysis toolset to quantify changes in complex organization. We applied the workflow to study the global protein complex rearrangements of THP-1 cells undergoing monocyte to macrophage differentiation and a subsequent stimulation of macrophage cells with lipopolysaccharide. We observed massive proteome organization in functions related to signaling, cell adhesion, and extracellular matrix during differentiation, and less pronounced changes in processes related to innate immune response induced by the macrophage stimulation. We therefore establish our integrated differential pipeline for rapid and state-specific profiling of protein complex organization with broad utility in complex experimental designs.

INSTRUMENT(S): TripleTOF 5600

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Culture

SUBMITTER: Ben Collins

LAB HEAD: Ben C. Collins

PROVIDER: PXD036711 | Pride | 2023-06-27

REPOSITORIES: Pride

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			Action	DRS
	20200720_080421_I181230_Report_helaccl2_SEC_long_Report_library.tsv	Tabular
	20200720_080421_I181230_Report_helaccl2_SEC_short_Report_library.tsv	Tabular
	SpecLib_cons_all.sptxt	Other
	aligned_filtered.csv	Csv
	iProphet.pep.xml	Pepxml

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Rapid profiling of protein complex re-organization in perturbed systems

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