Fig. 6: MncA read-outs of in-cell metalation in high Ni^II and Co^II refine metal availabilities.

a In-cell metalation with Ni^II and Co^II, not Cu^I or Zn^II, switches from Fe^II-MncA in metal-supplemented (600 µM, 300 µM, 600 µM, 800 µM respectively as shown) media, qualitatively matching predictions but quantitatively greater Ni^II and Co^II metalation than predicted. Metalation (dark blue bars) predicted using Supplementary Data 4 from availabilities in un-supplemented medium but with separately altered high availabilities of Cu^I, Zn^II, Ni^II or Co^II (as inset text). High availabilities correspond to the respective upper metal boundaries (θ_DM or θ_D, 0.99 or 0.01) shown in Supplementary Fig. 4. Mean ( ± SD) measurements of in-cell metalation (n = 3 independent biological replicates, square, circle, triangle) in high metal (pale blue bars) calculated using data in Supplementary Tables 6–9 based on Supplementary Figs. 11–15. b Selected intracellular metal availabilities (∆G_M) refined (triangles) using Supplementary Data 5 for Ni^II (left) from observed in-cell Fe^II- and Ni^II-MncA occupancies in high Ni^II (relative to Fe^II in un-supplemented medium), and for Fe^II (right) from Co^II and Fe^II occupancies in Co^II (relative to elevated Co^II as show in inset text). Metal preferences of MncA shown as ∆G_MP (circles). Bars are sensor ranges from 1% to 99%. c Calculated metalation (dark blue bars) using Supplementary Data 4 but with refined availabilities as in panel (b), reduced residuals for Mn^II (which was not included in the refinement process) relative to observed in-cell metalation (pale blue bars included for comparison), showing mean ( ± SD) (n = 3 independent biological replicates, square, circle, triangle). MncA metalation can be used to refine relative intracellular metal availabilities and Supplementary Data 5 is provided to enable such calculations. Source data are provided as a Source Data file.