SDS-PAGE analysis of purified App1p and the elution profile of PAP activity after chromatography with Superdex 200.
A, App1p was expressed and purified from S. cerevisiae as described under “Experimental Procedures.” Lane 1, molecular mass standards; lane 2, a sample (1.25 μg) of the purified App1p (indicated by the arrow). The SDS-polyacrylamide gel was stained with Coomassie Brilliant Blue R-250. B, purified App1p was subjected to Superdex 200 chromatography. Fractions (0.5 ml) were collected and assayed for PAP activity. The molecular mass standards (indicated by arrows from left to right) were β-amylase (200 kDa), alcohol dehydrogenase (150 kDa), bovine serum albumin (66 kDa), carbonic anhydrase (29 kDa), and cytochrome c (12.4 kDa).