ultraID: a compact and efficient enzyme for proximity-dependent biotinylation in living cells
Zhao, X.; Bitsch, S.; Kubitz, L.; Schmitt, K.; Deweid, L.; Roehrig, A.; Cappio Barazzone, E.; Valerius, O.; Kolmar, H.; Bethune, J.
Show abstract
Proximity-dependent biotinylation (PDB) combined with mass spectrometry analysis has established itself as a key technology to study protein-protein interactions in living cells. A widespread approach, BioID, uses an abortive variant of the E. coli BirA biotin protein ligase, a quite bulky enzyme with slow labeling kinetics. To improve PDB versatility and speed, various enzymes have been developed by different approaches. Here we present a novel small-size engineered enzyme: ultraID. We show its practical use to probe the interactome of Argonaute-2 after a 10 min labeling pulse and expression at physiological levels. Moreover, using ultraID, we provide a membrane-associated interactome of coatomer, the coat protein complex of COPI vesicles. To date, ultraID is the smallest and most efficient biotin ligase available for PDB and offers the possibility of investigating interactomes at a high temporal resolution.
Matching journals
The top 4 journals account for 50% of the predicted probability mass.
Similar papers in this journal
- The AAA+ ATPase RavA and its binding partner ViaA modulate E. coli aminoglycoside sensitivity through interaction with the inner membrane 97%
- Evolution of protease activation and specificity via alpha-2-macroglobulin-mediated covalent capture 97%
- Conformational coupling of the sialic acid TRAP transporter HiSiaQM with its substrate binding protein HiSiaP 96%
Similar papers in this journal
"Similar papers" are the closest papers from that journal in the model's embedding space. They show what the match is built on, but the ranking comes mostly from a classifier over the whole training set, not from these examples alone.