FIG. 5.

In-gel detection of MBH-dependent hydrogenase activity in the cytoplasmic fraction of TF140 (A) and the restoration of MBH targeting to the membrane by tat genes (B). (A) Lane 1, H16; lane 2, HF359, defective in the [NiFe]-containing subunit of MBH (ΔhoxG in pHG1); lane 3, TF140. (B) Analysis of restoration of MBH translocation was carried out by immunoblot analysis. S, the soluble extract consisting of the periplasmic and cytoplasmic fractions; M, the membrane fraction. MBH activity (milliunits per milligram of protein) is given below the blot. Lane 1, HF345, defective in the MBH-specific protease (ΔhoxM in pHG1); lane 2, TF140; lane 3, TF140 with tatAB (pGE600); lane 4, TF140 with tatA (pGE601); lane 5, TF140 with tatB (pGE602).