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. 2012 Oct 7;287(48):40670–40679. doi: 10.1074/jbc.M112.405704

FIGURE 4.

FIGURE 4.

Overexpression of Sdh5 partially restores growth of the single R638A mutant; the double C630A,R638A is unaffected. Similar effects are observed for steady-state covalent flavinylation and protein levels. A, growth test of strains with chromosomally integrated SDH1 C-terminal point mutants minus (top panel) or plus (bottom panel) low-copy vector (vec) expressing Sdh5. Cells were grown on glucose (fermentable) and glycerol/lactate (non-fermentable) media at 30 °C. B, upper panel: SDH activities of isolated mitochondria from strains in A expressed as a percentage of WT (n ≥ 3 ± S.D.). Lower panels: corresponding immunoblot analysis of mitochondria isolated from above strains showing steady-state levels of covalent flavinylation levels (UV illumination) and SDH subunit. The flavoprotein (Flavo) Δ% is a relative band density (quantified using ImageJ software) comparison of the Sdh5 overexpressing strain to that of the non-overexpressing strain.