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APEX is an engineered peroxidase that functions as an electron microscopy tag and a promiscuous labeling enzyme for live-cell proteomics. Because limited sensitivity precludes applications requiring low APEX expression, we used yeast-display evolution to improve its catalytic efficiency. APEX2 is far more active in cells, enabling the use of electron(More)
Neurexin and neuroligin are transmembrane adhesion proteins that play an important role in organizing the neuronal synaptic cleft. Our lab previously reported a method for imaging the trans-synaptic binding of neurexin and neuroligin called BLINC (Biotin Labeling of INtercellular Contacts). In BLINC, biotin ligase (BirA) is fused to one protein while its(More)
Here we discuss our additional efforts to understand why the A134P mutation in APEX2 renders it far more active than APEX inside cells. In the main text, we described how the most striking difference we could detect between APEX and APEX2 was their varying susceptibilities to H 2 O 2-induced inhibition. In the presence of 1.4 mM of the model phenolic(More)
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