Proteasome dysfunction during dopaminergic degeneration induces proteolytic stress, and is a contributing factor for the onset and formation of Lewy bodies. Results from our previous studies showed that synthetic prot...Proteasome dysfunction during dopaminergic degeneration induces proteolytic stress, and is a contributing factor for the onset and formation of Lewy bodies. Results from our previous studies showed that synthetic proteasome inhibitor-induced inclusions in PC12 cells contained six subunits in the 26S proteasome. In the present study, mass spectrometry analysis of single protein spots resolved by two-dimensional gel electrophoresis and identified by bioinformatic analysis of peptide mass fingerprint （PMF） data were performed to comprehensively characterize the proteomic profile of the proteasome subunits. Results showed that six subunits in the 26S proteasome were characterized through accurate assignment by PMF data-specific protein identification in protein databases. Additionally, identification of one of the proteasome subunits was further confirmed using a subunit-specific antibody against non-adenosine triphosphatase subunit 11 of the 19S regulatory particle. Results suggest that the potential proteomic profile of six subunits in the 26S proteasome could be established from proteasome inhibitor-induced inclusions in PC12 cells.展开更多
基金the Science and Technology Commission Foundation of Jilin Province,No.200505200the Distinguished Professor Foundation of Jilin University,No.450011011204
文摘Proteasome dysfunction during dopaminergic degeneration induces proteolytic stress, and is a contributing factor for the onset and formation of Lewy bodies. Results from our previous studies showed that synthetic proteasome inhibitor-induced inclusions in PC12 cells contained six subunits in the 26S proteasome. In the present study, mass spectrometry analysis of single protein spots resolved by two-dimensional gel electrophoresis and identified by bioinformatic analysis of peptide mass fingerprint （PMF） data were performed to comprehensively characterize the proteomic profile of the proteasome subunits. Results showed that six subunits in the 26S proteasome were characterized through accurate assignment by PMF data-specific protein identification in protein databases. Additionally, identification of one of the proteasome subunits was further confirmed using a subunit-specific antibody against non-adenosine triphosphatase subunit 11 of the 19S regulatory particle. Results suggest that the potential proteomic profile of six subunits in the 26S proteasome could be established from proteasome inhibitor-induced inclusions in PC12 cells.
文摘目的 用双向凝胶电泳-质谱法(2-DE/MS)分析人CD4+ CD25+T细胞和CD4+CD25-T细胞表达的蛋白质组,建立二者之间的差异表达谱。方法提取人脾脏源CD4+ CD25+T细胞和CD4+CD25-T细胞总蛋白质,双向电泳分离蛋白,比较两种细胞的蛋白质组表达差异,对差异蛋白质点用基质辅助激光解析电离飞行时间质谱(Matrix-assisted laser des-orption/ionization time of flying mass spectrometry,MALDI-TOF-MS)进行鉴定。结果胶图差异分析发现25个表达水平存在明显差异的蛋白质点,其中17个点在CD4+CD25-T细胞上调,8个点在CD4+CD25+T细胞上调。质谱初步鉴定出15个差异蛋白质点。结论CD4+ CD25+T细胞和CD4+ CD25-T细胞表达蛋白质组存在差异,这种蛋白质组的差异分析有助于进一步研究调节性T细胞功能相关蛋白质。
