changes in proteasome structure and function caused by hamlet in tumor cells蛋白酶体结构和功能的变化引起的哈姆雷特在肿瘤细胞.pdfVIP

Changes in Proteasome Structure and Function Caused by HAMLET in Tumor Cells 1 1 ¨ 2 1 1 Lotta Gustafsson , Sonja Aits , Patrik Onnerfjord , Maria Trulsson , Petter Storm , Catharina Svanborg1,3* 1 Institute of Laboratory Medicine, Department of Microbiology, Immunology and Glycobiology, Lund University, Lund, Sweden, 2 Department of Experimental Medical Science, Lund University, Lund, Sweden, 3 Singapore Immunology Network (SIgN), Biomedical Sciences Institutes, Agency for Science, Technology, and Research (A*STAR), IMMUNOS, BIOPOLIS, Singapore, Singapore Abstract Background: Proteasomes control the level of endogenous unfolded proteins by degrading them in the proteolytic core. Insufficient degradation due to altered protein structure or proteasome inhibition may trigger cell death. This study examined the proteasome response to HAMLET, a partially unfolded protein-lipid complex, which is internalized by tumor cells and triggers cell death. Methodology/Principal Findings: HAMLET bound directly to isolated 20S proteasomes in vitro and in tumor cells significant co-localization of HAMLET and 20S proteasomes was detected by confocal microscopy. This interaction was confirmed by co-immunoprecipitation from extracts of HAMLET-treated tumor cells. HAMLET resisted in vitro degradation by proteasomal enzymes and degradation by intact 20S proteasomes was slow compared to fatty acid-free, partially unfolded a- lactalbumin. After a brief activation, HAMLET inhibited proteasome activity in vitro and in parallel a change in proteasome structure occurred, with modifications of catalytic (b1 and b5) and structural subunits (a2, a3, a6 and b3). Proteasome inhibition was confirmed in extracts