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首页> 外文会议>American Society for Mass Spectrometry Conference on Mass Spectrometry and Allied Topics >Towards Analysis of Differentially Expressed Proteins in Drosophila Testes Using LTQ Orbitrap and Tandem Mass Tag
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Towards Analysis of Differentially Expressed Proteins in Drosophila Testes Using LTQ Orbitrap and Tandem Mass Tag

机译：使用LTQ orbitrap和串联质量标签分析果蝇睾丸中差异表达蛋白质的分析

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摘要

Mascot search identified a total of 438 proteins from the merged data file, of which 227 were considered significant matches (P < 0.05). Of the 227 proteins identified, 127 were successfully quantified, showing ratios in the two samples (127/126) 0.8approx1.0 for Drosophila pseudoobscura proteins and 0.2 for the standard BSA spiked in the samples. Fractionation of peptide mixture prior to LCMSMS analysis is essential to successful identification and quantitation of complex protein mixtures. A test analysis of the entire mixture with a 3-hour LCMSMS analysis only identified 70 proteins with 2 quantified, BSA was identified but not quantified successfully. pI based fractionation of peptides is proven to be efficient and effective. The relatively even distribution of peptides over the pH range reduces complexity of each fraction and thus allows identification of the maximum number of peptides in each fraction. Successful focusing of the majority of peptides (91.5percent) into one or two fractions maintains sensitivity for peptide analysis.
机译：吉祥物搜索鉴定了合并数据文件中总共438个蛋白质，其中227被认为是显着的比赛（P <0.05）。在鉴定的227个蛋白质中，成功地定量了127个，显示出两样样品（127/126）0.8appox1.0的比例为果蝇伪蛋白，0.2用于样品中的标准BSA。在LCMSMS分析之前的肽混合物的分馏对于成功鉴定和定量复杂蛋白质混合物的分馏是必不可少的。与3小时LCMSMS分析的整个混合物的测试分析仅鉴定了70种蛋白质，其中BSA被鉴定但未成功地定量。基于PI的肽分馏被证明是有效且有效的。在pH范围内的肽的相对均匀的分布降低了每个部分的复杂性，从而允许鉴定每个级分中的最大肽数。成功将大部分肽（91.5）聚焦到一个或两个级分中保持肽分析的敏感性。

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