FIG. 2.

In vitro interaction between the HupUV and HupT proteins. The proteins (40 pmol of HupUV and 120 pmol of HupT) were incubated for 15 min at 30°C in a buffer containing 20 mM Tris-HCl (pH 8), 0.15 M NaCl, and 10% glycerol. The proteins were then run on a native 7.5% acrylamide gel in 0.5× Laemmli buffer, and the gel was revealed either by hydrogenase activity staining (A) or with anti-His₆ tag antibodies (B). (C) Effect of H₂ on the HupUV-HupT interaction. HupUV was first incubated with H₂ during 15 min, and then HupT was added, and the incubation lasted for 15 additional min before the complexes were loaded on the gel.