Figure 4. Hsp12 causes membrane remodeling.

(A) DMPG liposomes (5 mg/ml) where incubated for 1 hr at room temperature in the presence or absence of Hsp12 (25 μg/ml) and/or in the presence or absence of trehalose (2% final). Pellet (p) and supernatant (s) fractions where separated by high-speed centrifugation, and lipid distribution was assessed by SDS-PAGE followed by altered Coomassie staining (10% acetic acid only). (B) DMPG liposomes (5 mg/ml) where incubated for 1 hr at room temperature in the presence or absence of Hsp12 (1.5–5 μg/ml). Pellet (p) and supernatant (s) fractions where separated by high-speed centrifugation, lipid distribution was assessed by SDS-PAGE followed by altered Coomassie staining (10% acetic acid only). (C) Samples for electron microscopy were prepared the same as in A, with Hsp12 at 2.5 μg/ml. Images where taken from samples prior to centrifugation. Samples where spread on glow-discharged EM grids and stained using 2% uranyl acetate.