Figure 1
From: Mapping the key residues of SufB and SufD essential for biosynthesis of iron-sulfur clusters
Effect of amino acid substitutions in SufB. (a) Growth phenotype of cells carrying mutations in sufB. The UT109 cells (Δisc Δsuf) harboring pUMV22 Spr were sequentially transformed with the pRK-sufCDSE plasmid and the pBBR-sufAB plasmid carrying a point mutation in sufB (SufBR226A, SufBN228A, SufBC254A, SufBQ285A, SufBW287A, SufBK303A, SufBC405A, or SufBE434A). The cells were grown on LB plates (without MVA) at 37 °C for 24 h. (b) Effect of amino acid substitutions on the maturation of [2Fe-2S] Fdx. Cells were grown in liquid Superbroth supplemented with glucose and MVA. Bacterial extracts were normalized with respect to cell density and subjected to native PAGE to separate holo- and apo-Fdx. Fdx was detected by Western blot analysis using specific antibody. (c) Protein levels of SufB, SufD, and SufC determined by SDS-PAGE and Western analysis.
