Figure 3

Cardiolipin efficiently reconstitutes NhaA dimers from monomers. Samples of monomeric NhaA suspension (12 µL, 1.7 µg protein, 7.6 µM NhaA) were prepared. One sample with no addition served as a control, and to the other samples different phospholipids were added at the indicated phospholipid/NhaA molar ratios: cardiolipin (CL) or L-a-Phosphatidylglycerol (PG) or L-a-phosphatidyl ethanolamine (PE). The phospholipid-containing samples were sonicated for 10 s in a bath sonicator at 23 °C and incubated for 45 min at 4 °C with slow agitation. Then, 15 µL of native gel sampling buffer was added, and the proteins were resolved on native gels. The experiment was conducted three times with identical results.