| In this research, both human wild type (WT) and mutant A4V SOD1 were encapsulated in wet-aged sol-gel glasses in an attempt to identify intermediates leading to aggregation. The encapsulated proteins were subjected to various conditions known to cause aggregation in solution such as heat, low pH ∼ 4.0, EDTA, TCEP or DTT, and TFE. Changes in the secondary structure of the entrapped SOD1 were monitored by using circular dichroism (CD) in the far-UV region.;Results showed major differences between the WT and A4V SOD1 secondary structures under most conditions. The encapsulation method is shown to enable long-term monitoring of the secondary structural changes over several months, and the optimal encapsulation method for the SOD1 is identified in order to minimize leaching. Finally, a possible common intermediate for both A4V and WT SOD1 was identified under different conditions. |
