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

Ubiquitin-Activated Interaction Traps (UBAITs) identify E3 ligase binding partners.

O'Connor HF, Lyon N, Leung JW, Agarwal P, Swaim CD, Miller KM, Huibregtse JM

We describe a new class of reagents for identifying substrates, adaptors, and regulators of HECT and RING E3s. UBAITs (Ubiquitin-Activated Interaction Traps) are E3-ubiquitin fusion proteins and, in an E1- and E2-dependent manner, the C-terminal ubiquitin moiety forms an amide linkage to proteins that interact with the E3, enabling covalent co-purification of the E3 with partner proteins. We designed UBAITs ... [more]

EMBO Rep. Dec. 01, 2015; 16(12);1699-712 [Pubmed: 26508657]

Throughput

  • Low Throughput

Additional Notes

  • The UBAITs (Ubiquitin-Activated Interaction Traps) method was used.

Curated By

  • BioGRID