On the Structure and Function of the Phytoene Desaturase CRTI from Pantoea ananatis, a Membrane-Peripheral and FAD-Dependent Oxidase/Isomerase
Figure 7
Effect of FAD on the formation of productive membrane associates.
A, SDS-PAGE of liposome-bound CRTI. Membrane binding was carried out in the presence (+FAD) or absence of FAD (-FAD). CRTI bound to liposomes was analyzed after two washing steps with buffer II (left traces) or after a buffer II and additional high-salt washing step (right traces). B, HPLC analysis of phytoene desaturation catalyzed by membrane-bound CRTI. Trace a; lycopene (2) formation from phytoene (1) by membrane-associated CRTI formed in the presence of FAD but incubated without subsequently adding free FAD (free FAD removed by the washing steps). Trace b, same experiment using CRTI associates prepared in the absence of FAD; the incubation contained 150 µM added FAD. Incubation time was 1 h at 37°C. HPLC trace represents a MaxPlot (250–550 nm).