FIG 5.
Introduction of the C-terminal peptide from HyaB onto pro-HybC delivers active Hyd-2 enzyme. (A) A native PAGE gel stained for hydrogenase activity using benzyl viologen-triphenyl tetrazolium chloride is shown. Samples of crude extracts (50 μg of protein) derived from MC4100 and FTD147 either lacking a plasmid or carrying a plasmid encoding pro-HyaBHybC or pro-HybCHyaB were loaded on the gel. (B) The same samples used for panel A were separated by 10% (wt/vol) SDS-PAGE and after transfer to nitrocellulose were challenged with anti-Hyd-2 antiserum. The migration positions of the respective polypeptides are indicated at the right. (C) The same samples used for panel A were stained for Hyd-1 enzyme activity with phenazine methosulfate-nitroblue tetrazolium dyes in the presence of H2.