质谱鉴定PC12细胞蛋白酶体抑制剂PSI-诱导性包含体中的LB蛋白质(英文)

Protein Components of Lewy Bodies(LBs) Identified from Synthetic Proteasome Inhibitor (PSI)-induced Inclusions in PC12 Cells by MS Analysis

路易(小)体(Lewy body,LB)构成特征的蛋白质组分在体外形成的路易(小)体样包含体(Lewy body-like inclusion)或聚集体(aggresome)中能够获得鉴定.通过蛋白质组学方法鉴定聚集体内蛋白质是一种新的途径.10μmol/L人工合成蛋白酶体抑制剂PSI(proteasomal inhibitor)作用PC12细胞48h,使其产生PSI诱导性包含体(PSI-induced inclusions).通过生物化学分级分离、双向电泳(two-dimensional electrophoresis,2-D)和肽质量指纹鉴定(identification via peptide mass fingerprints,PMF)的蛋白质组学方法,2个涉及突触递质合成的蛋白质、6个26S蛋白酶体亚基、2个细胞骨架蛋白、2个线粒体蛋白、1个抗氧化蛋白和7个分子伴侣蛋白和(或)分子伴侣样蛋白等20个LB蛋白质组分获得鉴定.结果提示,当PC12细胞发生蛋白酶体抑制时,这20个LB蛋白质组分可能被富集到PSI诱导性包含体中.

Proteomic analysis is an effective way to identify protein constituent in Lewy body-like inclusions （or aggresome） in vitro. Exposure to synthetic proteasome inhibitor （PSI, 10 μmol/L） for 48 hours was used to induce the formation of cytoplasmic proteineous inclusions （termed as PSi-induced inclusions） in PC12 cells. The proteomic approaches of biochemical fractionation, two-dimensional electrophoresis （2-D） and identification via peptide mass fingerprints （PMF） were deployed, and 20 protein components of LBs were identified, including 2 proteins involved in the production of synaptic neurotransmitter, 6 subunits of the 26 S proteasome, 2 cytoskeleton proteins, 2 subunits of mitochondrial complexes, 1 anti-oxidant protein, and 7 chaperone proteins and （or） chaperone-like proteins. The results suggested that these LB protein components might had been recruited in PSI-induced inclusions formed in PC12 cells under the condition of proteasome inhibition.