Figure 5

Protection of the binding site for I-concanolide A by plecomacrolidic antibiotics. Tricine-SDS-PAGE gels. Lane 1, stained with Coomassie Blue; lane 2–8, autoradiography of the gel after exposition to a phosphoscreen. Samples of 20 μg V-ATPase were preincubated for 60 min on ice with 100 μM or 10 μM bafilomycin B₁ (lanes 3 and 4), 100 μM or 10 μM apicularen A (lanes 5 and 6) and 100 μM or 10 μM archazolid (lanes 7 and 8), respectively. I-concanolide A was then added to give a final concentration of 10 μM. The mixture was incubated for another 60 min on ice and then treated with UV light. Control with pre-incubation, but without effectors (lane 2).