FIG. 1.

Immunoblot analysis of cross-linked subunit c (cA21C/cM65C) in inverted membrane vesicles of cells grown in the presence of different carbon sources. Membranes (5 mg/ml) prepared from E. coli DK8/pBWU13.NOC containing bi-cysteine-substituted subunit c were cross-linked with 1.5 mM copper-1,10-phenanthroline at pH 8.0, separated by nonreducing sodium dodecyl sulfate-polyacrylamide gel electrophoresis (20 μg/lane), and immunoblotted as described by Krebstakies et al. (17). Blot membranes were incubated with anti-c monoclonal antibody GDH 9-2A2 (2) as the primary and IRDye800DX-labeled goat anti-mouse immunoglobulin G (Rockland, Gilbertsville, PA) as the secondary antibody and finally detected by using the two-channel Odyssey system (LI-COR). MM, minimal medium.