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. 2005 Oct 25;391(Pt 3):567–574. doi: 10.1042/BJ20050718

Figure 4. HPLC and MS analyses of a GS assay with a non-natural co-substrate.

Figure 4

(A) HPLC fluorescence trace of S-bimane-derivatized reaction products of purified recombinant TaGS2 assay using γ-GC and D-alanine as substrates. Peak 1, excess γ-EC in the assay. Peak 2, novel reaction product (γ-Glu-Cys-D-Ala), which was not observed when TaGS2 was incubated with γ-EC minus D-alanine. Peak 3, bromobimane degradation product. (B) Control incubation of (A), with TaGS2 incubated with γ-GC only. (C) Upper and lower traces: mass spectral analysis of total reaction mixture shown in (A) and (B) respectively before derivatization with monobromobimane. The (M+H)+ at 251.1 corresponds to unchanged γ-GC, whereas the novel peak at 322.1 present in the upper trace but not in the lower trace corresponds to γ-Glu-L-Cys-D-Ala in trace (A).