Synthetic protease inhibitor-induced inclusions in PC12 cells Potential proteomic characterization of six subunits in the 26S proteasome  

作　　者：Mingxiu Tian Xing'an Li Yingjiu Zhang Yihong Hu Ming Chang Tao Liu Danping Wang Yu Zhang Linsen Hu 

机构地区：[1]Department of Neurology, First Affiliated Hospital of Jilin University, Changchun 130021, Jilin Province, China [2]Department of Neurology, Fourth Municipal Hospital of Tianjin, Tianjin 300140, China [3]Key Laboratory for Honeybee Genetics and Queen Breeding, Jilin Provincial Apiculture Institute, Jilin 132108, Jilin Province, China [4]Key Laboratory for Molecular Enzymology and Engineering, the Ministry of Education (Jilin University [5]College of Life Science, Jilin University, Changchun 130021, Jilin Province, China

出　　处：《Neural Regeneration Research》2010年第22期1685-1693,共9页中国神经再生研究（英文版）

基　　金：the Science and Technology Commission Foundation of Jilin Province,No.200505200;the Distinguished Professor Foundation of Jilin University,No.450011011204

摘　　要：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.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.

关 键 词：PC12 cells proteasome inhibitor-induced inclusions proteasome subunit two-dimensional gel peptide mass fingerprints PROTEOMIC 

分 类 号：Q511[生物学—生物化学] S332.3[农业科学—作物遗传育种]