Extended Data Fig. 8: Validation of metal binding in purified Desor_0198 (a), SVEN_5263 (b), CbrC (c) and CitX (d) by ICP-MS.
From: Co-evolution-based prediction of metal-binding sites in proteomes by machine learning
For a and b, the coevolved metal-binding cluster predicted by MetalNet are shown on the left, SDS-PAGE of the purified wild-type protein in the middle and ICP-MS measurement of metal binding of the purified protein are shown on the right. Both proteins are validated with zinc-binding activity. c, The coevolved metal-binding cluster predicted by MetalNet are shown on the left, SDS-PAGE of the purified wild-type protein, two single mutants (C56S or C182S) and the double mutant (C56S&C182S) in the middle and ICP-MS analysis of zinc binding in the purified wild-type cbrC and mutants on the right. Partial zinc binding was retained in each of the single mutant whereas metal binding activity was completely abolished in the double mutant. In a, b, and c, error bars mean and s.d (n = 3 biologically independent samples). ICP-MS analysis of Fe and Cu binding is measured only once. d, SDS-PAGE of the purified wild-type citX as well as four single mutants of the predicted metal-binding residues by MetalNet (C145S, C148S, C155S and H161S). ICP-MS analysis of citX is shown in Fig. 4d. The experiment was repeated twice independently with similar results.
