Dicyclohexylcarbodiimide inhibits proton pumping in ubiquinol:cytochrome c oxidoreductase of Rhodobacter sphaeroides and binds to aspartate-187 of cytochrome b.

In recent studies we reported that dicyclohexylcarbodiimide (DCCD) inhibited proton translocation in ubiquinol:cytochrome c oxidoreductase (cytochrome bc1 complex) from yeast mitochondria where it was bound to aspartate-160 of cytochrome b. In the current study, we report that DCCD and its fluorescent analogue, N-cyclohexyl-N'-[4-(dimethylamino)naphthyl]-carbodiimide (NCD-4), inhibit 50-60% proton pumping in the cytochrome bc1 complex of the bacterium Rhodobacter sphaeroides with a 20% inhibition of electron transfer activity. Radioactive DCCD is bound exclusively to cytochrome b at aspartate-187, which is located at the C-terminal region of the CD loop connecting membrane-spanning helices C and D of cytochrome b. Fluorescent studies with NCD-4 revealed that aspartate-187 is located in a mildly hydrophobic pocket in the bc1 complex at a distance of 2-3 A from the surface of the membrane.