摘要 |
We systematically investigated proteasome-mediated generation of fourteen different well-defined CTL epitopes. Synthetic peptides (26 residues) containing known CTL-epitopes flanked by their natural amino acids have been used as substrates for the 20S proteasome <i>in vitro</i>. After several time intervals, peptide digests were analyzed by electrospray mass spectrometry to determine the major fragments produced by the proteasome. In 12 out of 14 peptide digests, the correct C-terminal residue of the CTL-epitope was generated by proteasomal cleavage. The N-terminal residue of the epitope was generally not exactly defined by the proteasome. In most cases, fragments with the correct C-terminal residue were elongated several amino acids at the N-terminus. For two CTL-epitopes we found that their longer precursor peptides, as generated by the proteasome, correlated with efficient TAP translocation. For one CTL-epitope we found that a natural mutation directly flanking the C-terminal residue of the CTL-epitope precursor disrupted the specific C-terminal cleavage site and resulted in a non-functional cleavage product. This study indicates that proper CTL-epitope generation requires correct C-terminal cleavage by the proteasome, and allows N-terminal elongation of CTL-epitope precursor peptides.
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申请人 |
RIJKSUNIVERSITEIT TE LEIDEN;SEED CAPITAL INVESTMENTS (SCI) B.V.;OSSENDORP, FERRY;MELIEF, CORNELIS, JOHANNES, MARIA;OFFRINGA, RIENK;KESSLER, JOHAN, HERMAN |
发明人 |
OSSENDORP, FERRY;MELIEF, CORNELIS, JOHANNES, MARIA;OFFRINGA, RIENK;KESSLER, JOHAN, HERMAN |