Kurzfassung

Our project aims at the identification and prediction of cleavage site used by constitutive and immunoproteasomes in the presence of the 19S cap complex. We have established over the past years the methodology and specific skills to analyse and quantifiy peptide fragments generated in vitro by 20S proteasomes using entire, unmodified proteins as substrates (Nussbaum et al., 1998; Toes et al., 2001). We attempt now to extend our knowledge towards the detailed analysis of proteasomal cleavage...Our project aims at the identification and prediction of cleavage site used by constitutive and immunoproteasomes in the presence of the 19S cap complex. We have established over the past years the methodology and specific skills to analyse and quantifiy peptide fragments generated in vitro by 20S proteasomes using entire, unmodified proteins as substrates (Nussbaum et al., 1998; Toes et al., 2001). We attempt now to extend our knowledge towards the detailed analysis of proteasomal cleavage specificity by digesting entire proteins by 26S proteasomes containing constitutive or immuno β-subunits. This type of analysis will provide information whether or not the interaction of the 19S cap structure with 20S proteasomes changes the cleavage specificity and therefore the pool of peptides from which CTL epitopes are selected. In order to increase the number of potential substrate proteins to be degraded by 26S proteasomes we will identify additional proteins substrates.» weiterlesen» einklappen