Alkyne tyramide

Alkyne tyramide is a clickable ascorbate peroxidase 2 (APEX2) probe. Alkyne tyramide substantially improves APEX-labeling efficiency in intact yeast cells, as it is more cell wall-permeant than APEX2 substrate biotin-phenol (BP). Alkyne tyramide also facilitates the identification of APEX-labeling sites, allowing the unambiguous assignment of membrane topology of mitochondrial proteins[1].

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Yeast cells expressing Su9-APEX2 are incubated with each probe at 2.5 mM final concentration for half an hour, and the labeling reaction is initiated by the addition of 1 mM H2O2. Alkyne tyramide is the most reactive APEX2 substrate for labeling the yeast proteome[1]. APEX2-mediated Alkyne tyramide labeling in the yeast mitochondria is more effective at probe concentrations above 1 mM[1]. Alkyne tyramide enables proteomic profiling in the mitochondrial matrix of intact yeast cells with exceptionally high specificity (94%), and offers higher coverage than the traditional APEX2 substrate biotin-phenol (BP)[1].

[1]. Li Y, et al. A Clickable APEX Probe for Proximity-Dependent Proteomic Profiling in Yeast. Cell Chem Biol. 2020;27(7):858-865.e8.