Extended Data Figure 8: Relative adenylation activity of MoeZ and the MoeZ(Cys360Ala) mutant.
From: Co-opting sulphur-carrier proteins from primary metabolic pathways for 2-thiosugar biosynthesis
a, Reaction scheme for the MoeZ-catalysed adenylation activity assay. The adenylation activities of MoeZ and its Cys360Ala mutant were inferred using a colorimetric assay to monitor the production of AMP (indicated by a decrease in NADH at 340 nm) when MoeZ or its Cys360Ala mutant was co-incubated with a sulphur-carrier protein (MoaD2) in the presence of ATP, NaSH, adenylate kinase (AK), pyruvate kinase (PK) and lactate dehydrogenase (LDH). b, The relative adenylation activity of MoeZ (open circles) and its Cys360Ala mutant (filled circles), as well as a no MoeZ/MoeZ(Cys360Ala) control (open squares), was measured by the coupled enzyme assay, as described in a. Little difference in the decrease in absorption at 340 nm was observed between MoeZ and its Cys360Ala mutant (compared with the control with no MoeZ), suggesting that the mutation at Cys360 had little effect on the adenylation activity of MoeZ.
