Figure 5

The presence of oxidized residues does not explain the differential degradation of oxidized proteins by the four 20S proteasome subtypes. Assay comparing the fluorescence emitted following the degradation of oxidized and unoxidized FRET (Fluorescence resonance energy transfer)-peptides by the four proteasome subtypes. Oxidized and unoxidized precursor peptides were derived from (a) calmodulin, (b) hemoglobin, (c) NY-ESO1 and (d) LCMV nucleoprotein. Unoxidized precursor peptides contained a methionine residue which was replaced by methionine sulfoxide (Mox) in the corresponding oxidized precursor. These precursor peptides were incubated with the four proteasome subtypes, and their degradation was monitored by measuring fluorescence at 420 nm. RFU at 420 nm of each sample was reported to the RFU at 420 nm of the unoxidized precursor degraded by the SP. The absolute RFU values at 420 nm of the unoxidized precursors degraded by the four proteasome subtypes are indicated on the histogram. All values are mean + SEM of three independent experiments.