Skip to main content
. Author manuscript; available in PMC: 2020 Jul 1.
Published in final edited form as: Chembiochem. 2019 May 3;20(13):1659–1663. doi: 10.1002/cbic.201800817

Figure 4.

Figure 4.

Monitoring a proteolysis reaction using 5HTP-4CDZ/GFP quencher/fluorophore pair. A) Map of the plasmid for mammalian expression of SUMO-52-5HTP-EGFP. B) The strategy for monitoring the proteolysis reaction. C) ESI-MS analysis and D) fluorescence spectra of purified SUMO-52-5HTP-EGFP before and after modification with 4CDZ. E) Normalized fluorescence spectra of the proteolysis reaction of 4CDZ-labeled SUMO-52-5HTP-EGFP (by SUMO-star protease) over time show complete recovery of fluorescence after 90 minutes. F) SDS-PAGE analysis confirms complete proteolysis at this time. Intact SUMO-EGFP, as well as cleaved SUMO and EGFP bands are shown. The band marked with * corresponds to SUMO-star protease.