Analyses of the Large Subunit Histidine-Rich Motif Expose an Alternative Proton Transfer Pathway in [NiFe] Hydrogenases
Figure 4
Western-analysis of amino acid mutant strains.
Amino acids implicated in the proposed proton transfer pathway were replaced by site-directed mutagenesis. HynL was detected using Anti-HupL antibody, 25 µg of total proteins were applied both for the controls and for the mutant strains. GB112131: T. roseopersicina ΔhynSL, ΔhupSL, ΔhoxH1 strain, the negative control; GB112131+pTHOE5M: T. roseopersicina ΔhynSL, ΔhupSL, ΔhoxH1 strain+wild type HynSL expressing vector, pTHOE5M, the positive control. Mutated strains were the follows: R487I: T. roseopersicina MR487I strain, D103L: T. roseopersicina MD103L strain, H104A: T. roseopersicina MH104A strain.
