Figure 4
From: Modulation of MagR magnetic properties via iron–sulfur cluster binding
Three conserved cysteines play different roles in iron–sulfur binding in clMagR. (a, b) Chemical reconstitution-mediated iron–sulfur cluster assembly on apo-clMagRC60A monitored by UV–Vis absorption (a) and CD spectroscopies (b). The samples of spectra shown are as-isolated clMagRC60A (light orange) and chemically reconstituted clMagRC60A (chem re clMagRC60A, orange). (c, d) chemical reconstitution-mediated iron–sulfur cluster assembly on clMagRC124A monitored by UV–Vis absorption (c) and CD spectroscopies (d). The samples of spectra shown are as-isolated clMagRC124A (light purple) and chemically reconstituted clMagRC124A (chem re clMagRC124A, purple). (e, f) chemical reconstitution-mediated iron–sulfur cluster assembly on pigeon clMagRC126A monitored by UV–Vis absorption (e) and CD spectroscopies (f). The samples of spectra shown are as-isolated clMagRC126A (light blue) and chemically reconstituted clMagRC126A (chem re clMagRC126A, blue). SDS-PAGE results were shown in the right of corresponding UV–Vis spectra as inserts (a, c, e). The theoretical mass of the clMagRC60A monomer, clMagRC124A monomer and clMagRC126A monomer were 16.38 kDa. (g, h) The UV–Vis absorption (c) and CD spectra (d) of clMagRC60A obtained by mixing apo-clMagRC60A and holo-IscU which was recorded before the addition of apo-clMagRC60A (dotted orange lines) and after incubation with apo-clMagRC60A for 180 min (orange lines). Protein and reagent concentrations are described in the Experimental procedures.
