Affinity Capture-Western

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 identified by Western blot with a specific polyclonal antibody or second epitope tag. This category is also used if an interacting protein is visualized directly by dye stain or radioactivity. Note that this differs from any co-purification experiment involving affinity capture in that the co-purification experiment involves at least one extra purification step to get rid of potential contaminating proteins.

Publication

Down-regulation of active ACK1 is mediated by association with the E3 ubiquitin ligase Nedd4-2.

Chan W, Tian R, Lee YF, Sit ST, Lim L, Manser E

ACK1 (activated Cdc42-associated kinase 1) is a cytoplasmic tyrosine kinase implicated in trafficking through binding to epidermal growth factor (EGF) receptor and clathrin. Here, we have identified a new ACK1-binding partner, the E3 ubiquitin ligase Nedd4-2, which binds ACK1 via a conserved PPXY-containing region. We show that this motif also binds Nedd4-related proteins and several other WW domain-containing proteins, including ... [more]

J. Biol. Chem. Mar. 20, 2009; 284(12);8185-94 [Pubmed: 19144635]

Throughput

  • Low Throughput

Additional Notes

  • species of bait not clear

Curated By

  • BioGRID