4-Hydroxy-2-Nonenal-Modified Glyceraldehyde-3-Phosphate Dehydrogenase Is Degraded by Cathepsin G in Rat Neutrophils

Figure 3

The gel filtration pattern of activity of the GAPDH-degrading enzyme in the cell extracts from neutrophils. The cell extracts from neutrophils were fractionated by Sephacryl S-200 HR column chromatography equilibrated with 50 mM sodium acetate buffer containing 1 M NaCl (pH 4.0). Activity of the GAPDH-degrading enzyme was assessed by the amount of the decreased GAPDH level, which was analyzed by Western blotting using an anti-GAPDH mAb and digitized immunoblots. Myeloperoxidase activity was determined spectrophotometrically. Elastase and cathepsin G activities were measured using fluorometric substrates.