Figure 9
FSEC profiles of solubilized crude membranes. Membranes were isolated from yeast cells induced for hERG-TEV-GFP-His 8 production at 15°C for 48 hours. Membranes were solubilized at a concentration of 2 mg/ml in FC-12 at a 3:1 detergent to protein ratio without any supplement (A), supplemented with 5 mM KCl (B), supplemented with 5.1 mg/ml cholesteryl-hemisuccinate (C) or both (D), with cholesteryl-hemisuccinate and 1 μM Astemizole (E) or with cholesteryl-hemisuccinate without Astemizole (F) as described in Methods section. Solubilizations E and F were separated in presence of 1 μM Astemizole. Solubilized membrane proteins were separated on a Superose 6 10/300 GL column. Molecular weight markers (GE Healthcare Life Science) separated on the same column eluted as follows: Blue Dextran 2000, 2000 kDa at void volume 8 ml (marked with an asterisk), Thyroglobulin 669 kDa at 12.5 ml, Ferritin 440 kDa at 14.5 ml, Aldolase 158 kDa at 16.3 ml, Conalbumin 75 kDa at 17.3 ml, Ovalbumin 44 kDa at 17.6 ml.