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. Author manuscript; available in PMC: 2018 Jan 1.
Published in final edited form as: Methods Enzymol. 2016 Dec 7;584:229–253. doi: 10.1016/bs.mie.2016.10.025

Figure 1.

Figure 1

Schematic diagram of the inducible reconstitution and fluorogenic intramembrane protease assay. Pure rhomboid protease and FITC-TatA substrate in detergent micelles are mixed with liposomes at low pH to inactivate the protease during reconstitution. Detergent is removed from the protein/lipid/detergent complexes by means of dilution followed by ultracentrifugation. Resuspension of the proteoliposome pellet in neutral reaction buffer re-activates the protease. The FITC fluorophore on the reconstituted substrate is quenched by its proximity to the membrane lipids, allowing detection of a fluorogenic signal as evidence of proteolytic release of the FITC-labeled amino-terminus.