FIGURE 3.

Structural and flotillin-1-binding properties of GST-fused HNgb or ZNgb. A, electronic absorption spectra of the ferric (bold line), ferrous deoxy (fine line), and ferrous-CO (dotted line) forms of GST-fused HNgb or GST-fused ZNgb. The concentration of each recombinant protein was ∼5 μm on the basis of heme content. The Q bands from 500 to 600 nm are enlarged by a factor of 3 on the perpendicular axis. B, GST pull-down assays with Ngb and flotillin-1. Truncated or full-length flotillin-1 was incubated with GST-HNgb, GST-ZNgb, or GST in buffer (pH 8.0), washed, and analyzed by Western blotting with rabbit anti-flotillin-1 polyclonal antibody.