FIG. 7.

Hydrogenase activity comigrates with Ehb. A 7% Blue Native PAGE gel was prepared anaerobically and loaded with triplicate cell extracts of the wild-type S2 and the ΔehbN mutant S965 (60 μg of protein in each lane). Lane 1, S2; lane 2, S965. Standard proteins (5 μg of each), including albumin from bovine serum (66 kDa, monomer; 132 kDa, dimer), β-amylase (200 kDa), and apoferritin (443 kDa), were used as molecular mass markers. After electrophoresis, the gel was sliced into three pieces for silver staining (A), in-gel hydrogenase activity staining (B), and Western blotting with antibody targeting EhbN (C). The position of the Ehb complex (marked by arrows) is indicated by the band detected by Western blotting in lane 1 of panel C.