Purification of huPON1 variants from E. coli. (a) SDS-PAGE of purification from His6-MBP-PON1 fusions from the pHMT plasmid using NiNTA chromatography. (1) ΔN-huPON1; (2) ΔHDL-huPON1; (3) ΔN-ΔHDL-huPON1; (4) g2e6p-huPON1. (b) Blot of SDS-PAGE of PON1 variants as His6-MBP-PON1 fusions with HisProbe-HRP (Pierce), demonstrating that many of the smaller proteins bear the 6 × His tag and are likely proteolytic fragments. (1) ΔHDL-huPON1 lysate; (2) and (3), purified ΔHDL-huPON1; (4) cleaved MBP; (5) purified g2e6p-huPON1; (6) purified ΔN-huPON1. (c) SDS-PAGE of purified proteins from the MBP-PON1-His6 constructs in pET11a with coexpression of DnaK/DnaJ/GrpE from pKJE7 (Takara Bioscience). (1) huPON1; (2) ΔN-huPON1; (3) ΔHDL-huPON1; (4) ΔN-ΔHDL-huPON1; (5) g2e6p-huPON1; (6) ΔN-g2e6p-huPON1.
