Table 1 The basic properties and features of enzymatic tags developed for enzyme-catalyzed proximity labeling approaches based on APEX and BioID
From: In vivo interactome profiling by enzyme‐catalyzed proximity labeling
Enzyme | Enzyme activity | Year | Size (kDa) | Source | Mutations | Features |
|---|---|---|---|---|---|---|
APEX | Ascorbate peroxidase | 2012 | 28 | Pea | K14D, W41F, E112K | applicability for high-resolution EM tagging of mammalian organelles and specific proteins [36] |
APEX2 | Ascorbate peroxidase | 2015 | 28 | Soybean | K14D, W41F, E112K, A134P | more sensitive and active in cells than APEX for both protein imaging by EM and proteomic mapping[35]; APEX-seq for subcellular RNA detection [19] |
BioID | Biotin ligase | 2012 | 35 | E. coli | BirA-R118G | introduced as a useful screening tool for interacting and neighboring proteins in native cellular environment [9] |
BioID2 | Biotin ligase | 2016 | 27 | A. aeolicus | R40G | functionally comparable to BioID, but with more-selective targeting, less biotin supplementation requirement, and enhanced labeling efficiency [45] |
BASU | Biotin ligase | 2018 | 28 | B. Subtilis | Amino acids 1–65 deleted, R124G, E323S, G325R | faster kinetics, increased signal-to-noise ratio compared to BioID, enables direct detection of RNA-protein interactions [17] |
TurboID | Biotin ligase | 2018 | 35 | E. coli | Q65P, I87V, R118S, E140K, Q141R, A146Δ, S150G, L151P, V160A, T192A, K194I, M209V, M241T, S263P, I305V | generates detectable biotinylated materials for analysis within minutes; a superior methods for in vivo studies [32] |
miniTurbo | Biotin ligase | 2018 | 28 | E. coli | Amino acids 1–63 deleted, Q65P, I87V, R118S, E140K, Q141R, A146Δ, S150G, L151P, V160A, T192A, K194I, M209V, I305V | suggested to be less stable than TurboID, but with reduced interference with trafficking and function of fusion protein; preferable when a precisely defined labeling time is the priority [32] |