Figure 5.

Carbonylation of bean nodule Lb. A, Amino acid sequence of Lb in which the carbonylated residues found in vivo are marked with asterisks. B, Immunoblot of purified Lb after 6 h of MCO at different FeCl₃ and ascorbate (ASC) concentrations. CT, Control omitting ASC and FeCl₃; Ox1, 2.5 mm ASC, 10 μm FeCl₃; Ox2, 12.5 mm ASC, 50 μm FeCl₃; Ox3, 25 mm ASC, 100 μm FeCl₃; Ox4, 50 mm ASC, 200 μm FeCl₃; Ox5, 75 mm ASC, 300 μm FeCl₃. C, Immunoblot of purified Lb after 6 h of MCO at different FeCl₃, ASC, and H₂O₂ concentrations. Ox5, 75 mm ASC, 300 μm FeCl₃; Ox6, 75 mm ASC, 300 μm FeCl₃, 1 mm H₂O₂; Ox7, 75 mm ASC, 300 μm FeCl₃, 5 mm H₂O₂. Gels were loaded with 10 μg of protein per lane. D, Immunoblot of bean nodule extracts. Gels were loaded with 50 μg of protein per lane. For B to D, the apparent molecular masses (kD) of the monomer (M), dimer (D), and tetramer (T) are indicated. The anti-Lb antibody and the secondary antibody were used at dilutions of 1:1,000 and 1:40,000, respectively. Blots are representative of at least three independent experiments.