Co-fractionation/mass spectrometry to identify protein complexes
- Resource Type
- Authors
- Janelle C. Leggere; Claire D. McWhite; Kevin Drew; Ophelia Papoulas; Vy Dang; Edward M. Marcotte; Wisath Sae-Lee
- Source
- STAR Protocols
STAR Protocols, Vol 2, Iss 1, Pp 100370-(2021)
- Subject
- Proteomics
Chromatography
General Immunology and Microbiology
Proteome
Chemistry
Bioinformatics
General Neuroscience
Pipeline (computing)
Fractionation
Chemical Fractionation
Plants
Mass spectrometry
Cell Fractionation
General Biochemistry, Genetics and Molecular Biology
Epitope
Mass Spectrometry
Protocol
Animals
Humans
lcsh:Science (General)
Chromatography, Liquid
lcsh:Q1-390
- Language
- English
- ISSN
- 2666-1667
Summary Co-fractionation/mass spectrometry (CF/MS) is a flexible and powerful method to detect physical associations of proteins. CF/MS can be applied to any tissue or organism without the need for protein-specific antibodies or epitope tags. Here, we outline two alternate protocols for MS preparation of samples (containing low or high salt) and a computational pipeline (cfmsflow) that together allow the successful application of this approach. These protocols are based on CF/MS of over 16 diverse organisms including plants and animals. For complete details on the use and execution of this protocol, please refer to McWhite et al. (2020).
Graphical abstract
Highlights • Co-fractionation/mass spectrometry (CF/MS) detects native protein associations • Experimental methods for mass spec preparation of 96-well native fractions • Computational pipeline to generate protein interaction maps from CF/MS data
Co-fractionation/mass spectrometry (CF/MS) is a flexible and powerful method to detect physical associations of proteins. CF/MS can be applied to any tissue or organism without the need for protein-specific antibodies or epitope tags. Here, we outline two alternate protocols for MS preparation of samples (containing low or high salt) and a computational pipeline (cfmsflow) that together allow the successful application of this approach. These protocols are based on CF/MS of over 16 diverse organisms including plants and animals.