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

The deubiquitinase ubiquitin-specific protease 20 is a positive modulator of myocardial ?1-adrenergic receptor expression and signaling.

Yu SM, Jean-Charles PY, Abraham DM, Kaur S, Gareri C, Mao L, Rockman HA, Shenoy SK

Reversible ubiquitination of G protein-coupled receptors regulates their trafficking and signaling; whether deubiquitinases regulate myocardial ?1-adrenergic receptors (?1ARs) is unknown. We report that ubiquitin-specific protease 20 (USP20) deubiquitinates and attenuates lysosomal trafficking of the ?1AR. ?1AR-induced phosphorylation of USP20 Ser-333 by protein kinase A-? (PKA?) was required for optimal USP20-mediated regulation of ?1AR lysosomal trafficking. Both phosphomimetic (S333D) and phosphorylation-impaired ... [more]

J. Biol. Chem. Dec. 15, 2018; 294(7);2500-2518 [Pubmed: 30538132]

Throughput

  • Low Throughput

Curated By

  • BioGRID