Molecular modification for MHC class I and II antigen presentation

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HIV-1 Gag protein was expressed in HeLa cells and influenza virus HA in MDCK cells. When the protein degradation kinetics were analyzed by Western blotting, we found the t1/2 of Gag 8-9 hrs and that of HA slightly shorter. Gag and HA were more rapidly degradated by the proteasome pathway, when ubiquitin was added to their N-termini. Based on the N-end rule, when the N-terminal methionine of Gag after ubquitin cleavage was substituted with arginine, the Gag was more efficiently degradated by the ubquitin/proteasome pathway. In contrast, the addition of LAMP1 to Gag altered the major degradation pathway for Gag from the proteasome pathway to the lysosomal pathway.