Structure prediction and activity analysis of human heme oxygenase-1 and its mutant

Figure 1 Construction of expression vector pBHO-1(M).

Figure 2 Prime simulated structure of whHO-1 and △hHO-1. Red: heme; Black: His 25 and Ala 25. Ala 25 loses contacting with heme as His 25 does in the heme-binding pocket.

Figure 3 Changes of bond angle, dihedral angle and chemical bond.

Figure 4 Molecular surfaces and electrostatic potential of whHO-1 and △hHO-1. The electrostatic potential of active pocket is –1.800.

Figure 5 Western blotting of whHO-1 and △hHO-1 expressed in DH5α . Lanes 1, 2: Expression products of pBHO-1 and pBHO-1(M) in DH5α induced with IPTG; lane 3: Control; lanes 4, 5: Expression products of pBHO-1 and pBHO-1(M) in DH5α not treated with IPTG; lane 6: Marker.

Figure 6 Q-Sepharose Fast Flow column chromatography. A: pH 7.4 buffer; B: pH 8.4 buffer.

Figure 7 SDS-PAGE analysis of purified whHO-1 and △hHO-1. Lane 1: Marker; lane 2: whHO-1; lane 3: △hHO-1.