FIGURE 3.

The GRAM domain of GbpC binds to the plasma membrane. A, confocal fluorescent images were taken from gbpC-null cells expressing GRAM-GFP in the absence and presence of latrunculin A or GRAMG2378A-GFP. The results show that GRAM-GFP localizes independently of F-actin at the plasma membrane, whereas the mutant GRAMG2378A-GFP shows a cytoplasmatic distribution. B, PIP strips containing spots with various phospholipids were incubated with lysates from cells expressing GRAM-GFP or GRAMG2378A-GFP. Binding of these proteins to phospholipids was visualized using an anti-GFP antibody. For direct comparison, both proteins were assayed on PIP strips in parallel and exposed on film together, and input protein was similar for both proteins.