(2016). Nature Methods, 13(9) , 763-769. doi: 10.1038/nmeth.3935. Article Pubmed
Protein (state) | t1/2 (s) | Power | Light | Mode | In Cell | Fusion | ˚C |
---|---|---|---|---|---|---|---|
TDsmURFP | 190.0 | Arc-lamp | Widefield |
We chose TeAPCα (15 kD) [as a precursor for smURFP] because it lacked 29 amino acids common to other APCαs. Expression of TeAPCα with heme oxygenase-1 (HO-1) and phycocyanobilin–ferredoxin oxidoreductase (PcyA) for PCB production showed no fluorescence.
In our approach, we first evolved TeAPCα mutants to autocatalytically attach PCB (without a lyase) and fluoresce. Second, since PCB is not present in mammals, we evolved derivatives that bind BV, a molecule ubiquitous in eukaryotes and produced at the rate of 300–500 mg per d in humans.