Fig. 3: SPA and LM-2I target ASS1 at C97 site. | Nature Communications

Fig. 3: SPA and LM-2I target ASS1 at C97 site.

From: Naturally-occurring spinosyn A and its derivatives function as argininosuccinate synthase activator and tumor inhibitor

Fig. 3

a The recombinant ASS1 protein was incubated with Biotin-SPA in the absence or presence of SPA, SPAH and LM-2I for 1 h at room temperature, followed by western blot using streptavidin-HRP or ASS1 antibody (n = 3 biologically independent experiments). b Recombinant WT and mutant ASS1 proteins were incubated with Biotin-SPA or Biotin-SPAH for 30 min, followed by western blot using streptavidin-HRP or ASS1 antibody (n = 3 biologically independent experiments). MS/MS analysis for the C97-containing peptide of recombinant ASS1 pre-incubated with vehicle (c), SPA (d), or LM-2I (e). (SPA), combined with SPA; (LM-2I), combined with LM-2I; (c), carbamidomethylation; (d), deamidation. f Binding model of LM-2I-ASS1. The left panel is the global view of the entire ASS1 structure, the right panel is the focused view of LM-2I in the binding site (ASS1 in gray while LM-2I in green). Source data are provided as a Source Data file.

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