Affinity Capture-MS

An interaction is inferred when a bait protein is affinity captured from cell extracts by either polyclonal antibody or epitope tag and the associated interaction partner is identified by mass spectrometric methods.

Publication

Composition and Regulation of the Cellular Repertoire of SCF Ubiquitin Ligases.

Reitsma JM, Liu X, Reichermeier KM, Moradian A, Sweredoski MJ, Hess S, Deshaies RJ

SCF (Skp1-Cullin-F-box) ubiquitin ligases comprise several dozen modular enzymes that have diverse roles in biological regulation. SCF enzymes share a common catalytic core containing Cul1â‹…Rbx1, which is directed toward different substrates by a variable substrate receptor (SR) module comprising 1 of 69 F-box proteins bound to Skp1. Despite the broad cellular impact of SCF enzymes, important questions remain about the ... [more]

Cell Nov. 30, 2017; 171(6);1326-1339.e14 [Pubmed: 29103612]

Throughput

  • Low Throughput

Additional Notes

  • SILAC-SRM (stable isotope labeling by amino acids in cell culture-based selected reaction monitoring) was used to determine percent exchange of isotopically heavy for light hit protein in immunoprecipitates of endogenous Cul1 under a variety of experimental conditions. From this it is assumed that an interaction with endogenous Cul1 was detected.

Curated By

  • BioGRID