Effect and mechanism of β-L-D4A on DNA polymerase α

Figure 1 A280 and gradient concentration of KPO₄ in each chromatography. The curve with spots is for A280, the other is for KPO₄. A: A broad peak of protein from the 16^th tube (0.115 mol/L) to the 90^th tube (0.185 mol/L), clearly observed in the second DE52 chromatography; B: Phosphocellulose chromatography displaying a major peak of protein identified at 0.165 mol/L KPO₄ and a minor trailing shoulder at 0.21 mol/L; C: Hydroxylapatite chromatography showing a single sharp peak of protein at 0.085 mol/L KPO₄; D: DNA-cellulose chromatography revealing a single sharp peak at 0.085 mol/L KPO₄.

Figure 2 SDS-PAGE displaying decreased bands after purification steps-DE52 (lane 1), P11 (lane 2), HTP (lane 3) and DNA (lane4), and M (marker), While DNA-cellulose chromatography demonstrating only one band (M_r 160 000).

Figure 3 Western blot of the fractions of DNA-cellulose chromatography. When mouse anti-human DNA polymerase α monocloned antibody was used as the primary antibody, the fraction was found to be DNA polymerase α.

Figure 4 Determination of detailed kinetic parameters Km (A), IC50 (B), and Ki (C for 3TC, D for β-L-D4A).