FIG. 5.
RH protein stability and RH-mediated H2-oxidizing activity in the hyp-negative strain HF581. R. eutropha strains were grown in FGN medium supplemented with or without 1 μM ZnCl2 as indicated above the figures. (A) Immunoblot against the RH large subunit HoxC, with 20 μg of soluble protein in each lane. (B) Immunoblot against the RH small subunit HoxB, with 20 μg of soluble protein in each lane. (C) In-gel activity assay. A total of 500 μg of soluble proteins were separated by native PAGE. Dark-colored bands indicate H2-oxidizing activity of the RH by the PMS-mediated reduction of NBT. The fact that the RH forms an α2β2 oligomer was previously described (2). Lanes 1 and 2, HF573; lanes 3 and 4, HF581.