Figure 3.

Eukaryotic rhomboid proteases and pseudopreoteases are monomeric in membranes. (A) Gel electrophoretic analysis of liposome-reconstituted NHalo-DmRho4 activity in the absence or presence of calcium. Shown is a fluorescence scan to reveal FITC-TatA-TM (lower band labeled in red denotes cleaved product). (B) Quantification of NHalo-DmRho4 photobleaching steps in planar bilayers in the presence or absence of calcium. (C) Quantification of photobleaching step events for six rhomboid proteins in the plasma membrane of HEK293T cells. A representative single-molecule TIRF microscopy image of RHBDL2-HaloC labeled with HTL-JF646 in HEK293T cells is shown on the right. To see this figure in color, go online.