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

Figure 5. HPLC analysis of wheat hmGSH and TaGS2 D-serine reaction product.

Figure 5

(A) HPLC trace of γ-glutamyltranspeptidase digest, run on a Chirobiotic V column, of the purified product of TaGS2 D-serine assay. MS identified peak 1 as undigested parent compound (S-bimane-derivatized γ-Glu-Cys-Ser: m/z+ 528.18), peak 2 as the S-bimane-derivatized Cys-Ser dipeptide product of the digest (m/z+ 399.13) and peak 3 as S-bimane-derivatized cys-teine (m/z+ 312.10). (B) HPLC trace of γ-glutamyltranspeptidase digest, run on a Chirobiotic V column, of hmGSH purified from wheat. MS confirmed the identity of each peak to be of identical mass to that shown in (A). (C) HPLC trace of purified peak 2 in (A), co-injected with S-bimane-derivatized L-Cys-D-Ser standard, on a Kingsorb C18 column. (D) HPLC trace of purified peak 2 in (B), co-injected with S-bimane-derivatized L-Cys-L-Ser standard, on a Kingsorb C18 column. (E) HPLC trace of S-bimane-derivatized L-Cys-L-Ser standard and S-bimane-derivatized L-Cys-D-Ser standard co-injected on to a Kingsorb C18 column. Peak 1 is S-bimane-derivatized L-Cys-L-Ser and peak 2 is S-bimane-derivatized L-Cys-D-Ser.