Figure 1.

Gel filtration column chromatography on Superose 12 FPLC. The active fractions obtained from Mono Q FPLC column were applied to a Superose 12 gel filtration FPLC column as described in “Materials and Methods.” The inset shows the calibration curve for the estimation of the apparent molecular mass of the PLA₂. The standard protein markers for the gel filtration chromatography were as follows: β-amylase (200 kD), alcohol dehydrogenase (150 kD), BSA (66 kD), carbonic anhydrase (29 kD), and cytochrome c (12.4 kD) (A). The active fractions from Superose 12 gel filtration FPLC columns were analyzed by 12% (w/v) SDS-PAGE and visualized by silver stain as described in “Materials and Methods.” The standard protein markers were as follows: myosin (200 kD), β-galactosidase (116 kD), phosphorylase b (97.4 kD), BSA (66.2 kD), ovalbumin (45 kD), carbonic anhydrase (31 kD), and trypsin inhibitor (21.5 kD). The molecular mass (arrow) of the plant PLA₂ was extrapolated from R_f value (B). Essentially identical results were obtained in three independent experiments.