Mass spectrometric analysis of Podospora anserina mitochondria

Here we report data of an analysis to monitor the age-related quantitative protein composition of the mitochondria of the fungal aging model Podospora anserina. The impact of senescence on mitochondrial protein composition was analyzed by LC-MS analysis. In an untargeted proteomic approach, we identified 795 proteins in samples from juvenile and senescent wild-type cultures and obtained quantitative information for 226 of these proteins by spectral counting. Despite the broad coverage of the proteome, no substantial changes linked to the aging process could be observed. LC-MS/MS-data were analyzed using the Sequest algorithm implemented in the Bioworks 3.3.1 software (Thermo scientific) for peptide identification versus a P. anserina protein database. As enzyme specificity Trypsin was selected. Two missed cleavages were permitted and the mass tolerance was set to 10 ppm. False discovery rates were estimated by the number of spectral matches to a decoy database. Acceptance criteria and filters were set to achieve a false positive rate of 5 %.

本研究报道了针对真菌衰老模型柄孢壳菌（Podospora anserina）线粒体的年龄相关蛋白质定量组成的分析数据。本研究采用液相色谱-质谱联用（LC-MS）技术，分析了衰老对线粒体蛋白质组成的影响。通过非靶向蛋白质组学方法，我们在幼龄与衰老野生型培养物的样本中鉴定出795种蛋白质，并通过光谱计数法获得了其中226种蛋白质的定量信息。尽管该蛋白质组覆盖范围较广，但未观察到与衰老过程相关的显著变化。
针对液相色谱-串联质谱（LC-MS/MS）数据，我们采用赛默飞世尔科技（Thermo scientific）Bioworks 3.3.1软件中集成的Sequest算法，以柄孢壳菌（P. anserina）蛋白质数据库为参照进行肽段鉴定。实验选择胰蛋白酶作为酶切特异性酶，允许最多2处漏切，质量容差设置为10 ppm。假发现率通过肽段谱图与诱饵数据库的匹配数进行估算，并设置鉴定标准与过滤参数，将假阳性率控制在5%以内。