Figure 3

Binding of pFNR and mutants to liposomes. (A) 5 mM (lipids) of liposomes with a diameter of 200 nm generated as described [41] and composed of PC (black bar), 85 mol% PC and 15 mol% DGDG (grey bar) or 85 mol% PC and 15 mol% PG (white bar), were incubated with labelled precursor and binding analyzed as described. The binding is expressed as percent of the added precursor. Shown is the average of at least three independent measurements. (B) Labeled pFNR (top) and pFNR F→G (bottom) was incubated with 0.1 mM (lane 1, 4, 7), 1 mM (lane 2, 5, 8) and 5 mM (lane 3, 6, 9) liposomes composed of PC (lane 1–3), 85 mol% PC and 15 mol% DGDG (lane 4–6) or 85 mol% PC and 15 mol% PG (lane 7–9) and binding determined as described, Lane 10 represents radioactive protein processed in the absence of liposomes. (C, D) Experiments as shown in B) were quantified and binding to liposomes composed of PC (black circle), 15 mol% DGDG/85 mol% PC (grey triangle) or 15 mol% PG/85 mol% PC (white diamonds) are expressed as ratio to input. The average of at least three independent measurements is expressed.