Table 2.
Distribution of PPK and PAP activity
| Fraction* | Protein (mg) | PPK Activity (units) | PAP Activity (units) |
|---|---|---|---|
| Before fractionation | 87.5 | 5.34 | 2.63 |
| I | 2.85 | <0.05 | <0.005 |
| II | 0.363 | <0.05 | <0.005 |
| III | 0.638 | <0.05 | <0.005 |
| IV | 17.0 | <0.05 | <0.005 |
| V | 25.0 | <0.05 | <0.005 |
| VI | 21.0 | <0.05 | <0.005 |
| VII | 9.50 | <0.05 | <0.005 |
| VIII | 11.3 | 0.930 | <0.005 |
| IX | 7.13 | 0.990 | <0.005 |
| X | 5.63 | 1.07 | <0.005 |
| XI | 0.538 | <0.05 | <0.005 |
*
The cell lysate containing overproduced PPK (87.5 mg of proteins) was put onto a DEAE-Toyopearl 650M column (1.6 × 12.5 cm; Toso) equilibrated with buffer P, followed by stepwise elution with buffer P containing 0, 50, 100, 150, 200, 250, 300, 350, 400, 450, and 500 mM NaCl. The eluents from each step were defined as fractions I to XI, respectively. Each fraction was subjected to the PPK and the PAP assays.