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The cysteine codon at position 1238 was mutated to valine in the context of full-length or truncated MEKK1. The proteins were overexpressed and then treated with NEM either in cells (A) (full-length, 0.25, 0.1 and 0.05 mM; truncations, 1 and 0.5 mM) or following purification (B) (1 mM NEM for all). The C1238V mutant of MEKK1 is completely resistant to NEM. Abbreviations: autoP, autophosphorylation; WT, wild-type; CV, C1238V mutant.
