Fig. 3: Synthesis and characterization of FP catenanes.

a Two-step synthesis of GFP catenanes via a pseudorotaxane intermediate. Two rings constituting cat-GFP are colored red and green, respectively. b SDS-PAGE analysis of rtx-GFP (left lane) and the reaction mixture (right lane), showing the yields of in vitro synthesis (~46% for [2]cat-GFP and ~37% for [3]cat-GFP as determined by gel densitometry). c SEC overlay of rtx-GFP (black), the reaction mixture (blue), purified [2]cat-GFP (green) and [3]cat-GFP (red). d Cartoon illustration of corresponding TEV protease digestion products. e SDS-PAGE analysis of the TEV protease digestion products. f Structure of [2]cat-GFP predicted by AF2. Two rings constituting [2]cat-GFP are colored red and green, respectively. g Structure of [3]cat-GFP predicted by AF2. Two rings constituting [3]cat-GFP are colored red and green, respectively. h LC-MS spectra of [2]cat-GFP and [3]cat-GFP. i LC-MS spectra of cat-mCherry (Two rings constituting cat-mCherry are colored red and pink, respectively), cat-mWasabi (Two rings constituting cat-mWasabi are colored red and cyan, respectively), and cat-YPet (Two rings constituting cat-YPet are colored red and yellow, respectively). Source data are provided as a Source Data file.