Fig. 1.
In vitro characterization of VidaL. (A) Schematic for protein trans-splicing reactions with canonically split (Top; in red) and atypically split (Bottom; in blue) inteins. Three N- and C-extein residues on either side of the splice junction are shown in green. (B) Schematic for in vitro PTS with VidaL to generate labeled GFP (Left) and a representative Western blot of the reaction (Right). FLAG-tagged starting material (in red) decreases over time, and spliced product (dually tagged) increases. (C) ESI mass spectrum (deconvoluted inset) for biotin-GFP-FLAG spliced product. (D) PTS half-lives across a range of temperatures (Left) and NaCl concentrations (Right); reaction as in B. Conditions: 2 μM VidC-GFP-FLAG, 4 μM biotin-VidN, splicing buffer (pH 7.2). Error bar indicates SD (n = 3).