高分辨飞行时间质谱在蛋白质组学相对定量分析中的应用

发布时间：2018-04-30 01:15

本文选题：定量蛋白质组学 + 高分辨飞行时间质谱　；参考：《分析化学》2016年03期

【摘要】：利用Triple TOF 5600高分辨质谱仪分析牛血清白蛋白等3种蛋白质标准品,研究了质谱离子强度与蛋白质样品相对含量的相关性。蛋白质标准品用胰酶酶切后,稀释成1~1024 fmol/7μL的系列溶液,考察在1~1024 fmol上样量情况下,肽段的前体离子计数(cps)、蛋白质全部肽段的离子计数之和以及被检出肽段数目与上样量的相关性,以及相同样品在3次平行实验之间这些数值的变化幅度。结果表明。被检出肽段数目与上样量正相关,当cps超过1000时,所有肽段离子强度之和与上样量呈线性关系,但是用最灵敏肽段的离子强度表示更为准确。3次测量同一肽段的最高离子强度通常不会超过最低强度的1.5倍,提示当不同样品中同一蛋白的离子强度相差3倍以上是判断不同样品中相同蛋白质的含量具有差异的可靠阈值。本研究提供了一种利用高分辨率和高扫描速度蛋白质组组学定性数据进行半定量分析的方法,简便、快速,可为相关生物学和医学研究提供参考。
[Abstract]:Three kinds of protein standard were analyzed by Triple TOF 5600 high resolution mass spectrometer, and the correlation between ion strength and relative content of protein samples was studied. After digested with trypsin, the standard protein was diluted into a series of solution of 1t0 024 fmol/7 渭 L, and the sample amount of 1 1 024 fmol was investigated. The number of precursors of the peptide segment is counted by the number of CPS, the sum of the ion counts of the whole peptide segment of the protein, the correlation between the number of the detected peptide segments and the amount of the sample, and the variation of these values between the three parallel experiments of the same sample. The results show that. The number of peptides detected was positively correlated with the amount of sample. When cps exceeded 1000, the sum of ionic strength of all peptide segments was linearly related to the amount of sample. But the ion strength of the most sensitive peptide segment is expressed more accurately by measuring the maximum ionic strength of the same peptide segment for more than 1.5 times the minimum intensity. It is suggested that the difference of ionic strength of the same protein in different samples is more than 3 times, which is the reliable threshold to judge the difference of the content of the same protein in different samples. This study provides a method of semi-quantitative analysis based on high resolution and high scanning speed qualitative data of proteomics. It is simple and rapid and can be used as a reference for related biological and medical research.
【作者单位】：深圳大学生命科学学院微生物工程重点实验室;深圳大学生命与海洋科学学院深圳市海洋生物资源与生态环境重点实验室;
【基金】：深圳市科技项目(Nos.JSGG20140703163838793,20150006)资助
【分类号】：Q503;O657.63

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