Figure 1
Stability and inhibitory activity of conserpin-AATRCL. (A) RCL sequence alignment indicating which residues of conserpin were replaced with the corresponding residues in α1-AT; (B) Variable temperature thermal melt of conserpin-AATRCL, heating to 95 °C (black line) and cooling to 35 °C (red line), measured by CD at 222 nm; (C) Spectral scan before (black line) and after (red line) variable temperature thermal melt; (D) Variable temperature thermal melt in the presence in 2 M GdnHCl (heating to 95 °C; black line, cooling: red line); (E) Inhibitory activity assay and (F) SI against trypsin (n = 3); (G) A cropped SDS-PAGE showing a serpin:protease complex formed between HNE and AAT, but less complex formed between HNE and conserpin-AATRCL. From left to right: 1. Molecular weight markers (kDa); 2. α1-AT alone; 3. 1:1 ratio of α1-AT: HNE; 4. 2:1 ratio of α1-AT:HNE; 5. HNE alone; 6. conserpin-AATRCL alone; 7. 1:1 ratio of conserpin-AATRCL:HNE; 8. 2:1 ratio of conserpin-AATRCL:HNE. The full length SDS-PAGE gel is presented in Fig. S1.
