Fig. 1.
Interaction of hGBP1 with GUVs. (A) GUVs labeled with 1% TMR-PIP2 (red) in the presence of 2.5 µM AF-hGBP1F (green) and 400 µM corresponding nucleotides after 5 min of incubation. (B) Same as in A, but in the presence of AF-hGBP1NF. (C) Mean fluorescence intensities of AF-hGBP1 binding to GUVs (Methods) for different nucleotides. Statistics include the data for n = 25–80 GUVs under each experimental condition from at least two independent measurements. (D) Time-dependent dissociation of hGBP1F from a GUV in the presence of GTP monitored on a single vesicle. The experiment was started by addition of 1 mM GTP (t = 0 s) into a solution containing 2.5 µM AF-hGBP1F. (E) Numerical representation of D. (F) Time-lapse images of AF-hGBP1F dissociation from GUVs in the presence of 400 µM GTPγS (Upper) or 400 µM GTP (Lower) (see also Movie S1). (G) Numerical representation of F. (Scale bars, 5 µm.) a.u., arbitrary units.