Skip to main content
. 2010 Nov 10;286(4):2795–2806. doi: 10.1074/jbc.M110.186999

FIGURE 4.

FIGURE 4.

Copper and zinc concentrations from detergent-insoluble and -soluble species suggest minimal SOD1 metallation in detergent-extracted SOD1. A, the detergent-insoluble species were prepared from nontransgenic and G93A, G37R, H46R/H48, and hWT SOD1 transgenic mice. SOD1 concentration was measured by densitometry from a Western blot, and the copper and zinc concentrations were measured by ICP-MS. Despite the variable levels of SOD1 from each detergent insoluble pellet preparation, the levels of copper and zinc did not fluctuate across all the mice types. B, detergent-insoluble and detergent-soluble species were analyzed for activity and SOD1 concentration. The relative activity was divided by the SOD1 concentration to gain a rough idea of the level of copper bound to the SOD1. The axis on the left is in arbitrary units. The soluble hWT, H46R/H48Q, and G37R SOD1s were measured as 2123, 172, and 1682, respectively, which was significantly higher activity per dimer than their detergent-insoluble counterparts, which measured as 143, 9, and 18, respectively. Note that very little WT hSOD1 was found in the detergent-resistant pellets. The box plots represent median values and 25th and 75th percentile ranges, and bar graphs represent mean ± S.D. of three or more independent mice per genotype.