Figure 4: The unstructured form of apo-G93A is fully reduced in the cell. | Nature Communications

Figure 4: The unstructured form of apo-G93A is fully reduced in the cell.

From: In-cell NMR reveals potential precursor of toxic species from SOD1 fALS mutants

Figure 4: The unstructured form of apo-G93A is fully reduced in the cell.

(a) Thiol alkylation reaction with AMS on cell extracts containing [13C,15N]Cys-labelled G93A and WT SOD1 expressed in excess of zinc was run on a non-reducing SDS–PAGE and detected with western blot analysis. The band shift corresponds to four linked AMS, indicating that all four cysteines were reduced. (b) 2D projections of a three-dimensional CBCANH NMR experiment, showing that the Cβ chemical shift of three out of four cysteines could be measured, and corresponds to that of reduced cysteines (CβSH), while no signal is detected in the region of the Cβ of oxidized cysteines (CβSS). Cα, CβSS and CβSH regions are showed as light grey areas. The crosspeaks arising from [13C,15N]Cys-labelled glutathione are marked with an asterisk.

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