Figure 4
Limited proteolysis to examine the effects of AHSP and β globin on folding of nascent α globin. (A) 35S-radiolabeled α globins were synthesized by TNT with or without recombinant AHSP (4 μg/mL) and then treated with 10 μg/mL trypsin for the indicated times. Digestion was stopped by addition of SDS-PAGE loading dye. The extent of proteolysis was determined by SDS-PAGE electrophoresis followed by autoradiography. (B) 35S-radiolabeled α globin was synthesized by TNT with 40 ng/mL β globin (2.5 nM) present and then treated with 15 μg/mL trypsin for the indicated times and processed as described in panel A. CN-heme was not added to the TNT reactions in these experiments.
