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

引用本文: 洪晓愉, 王浩, 徐金玲, 李水明, 王勇. 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[J]. 分析化学, 2016, 44(3): 403-408. doi: 10.11895/j.issn.0253-3820.150751 shu

Citation: HONG Xiao-Yu, WANG Hao, XU Jin-Ling, LI Shui-Ming, WANG Yong. Application of High Resolution Time-of-Flight Mass Spectroscopy in Relative Quantitative Analysis in Proteomics[J]. Chinese Journal of Analytical Chemistry, 2016, 44(3): 403-408. doi: 10.11895/j.issn.0253-3820.150751 shu

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

洪晓愉 ^1, ,
王浩 ^1, ,
徐金玲 ^1, ,
李水明 ^2, ,
王勇 ^{2,
,}

1.
¹ 深圳大学生命科学学院, 微生物工程重点实验室, 深圳 518060;
2.
² 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, 深圳 518060

通讯作者: 王勇

基金项目:
本文系深圳市科技项目(Nos.JSGG20140703163838793,20150006)资助 (Nos.JSGG20140703163838793,20150006)

摘要: 利用TripleTOF 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/7 μL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。

关键词:

English

Application of High Resolution Time-of-Flight Mass Spectroscopy in Relative Quantitative Analysis in Proteomics

1.
¹ College of Life Science, Shenzhen Key Laboratory of Microorganism, Shenzhen University, Shenzhen 518060, China;
2.
² College of Life Science, Shenzhen Key Laboratory of Marine Bioresources and Ecology, Shenzhen University, Shenzhen 518060, China

Corresponding author: 王勇

Received Date: 22 September 2015
Fund Project:
本文系深圳市科技项目(Nos.JSGG20140703163838793,20150006)资助

Abstract: By using the high resolution mass spectrometer TripleTOF 5600, three kinds of standard proteins including bovine serum albumin (BSA), ovalbumin (OVA) and lysozyme C(LYZC) were analyzed, and the correlationship between the ion intensity of mass spectrometry and the relative content of protein sample was investigated. The protein samples were digested by trypsion and diluted to 1-1024 fmol in 7 μL. The ion counts per second (cps) were used to stand for the amounts of proteins and peptides. Then the correlation between sum of ion intensity (cps) of all the peptides, number of peptides detected and the amount of proteins was investigated. By comparing the change of values of the same sample in three parallel experiments, a linear relationship between these indexes and the amount of proteins within 1-1024 fmol was found when the cps was more than 1000. Usually, the maximal ion intensity was no more than 1.5 times of the minimum value for same peptide in triplicate experiments, which suggested that the 3 times or more change of ion intensity was the minimum threshold to determine the differences of proteins amounts in different samples. This study provides a relative quantitative analysis method using qualitative data of high resolution and high scan speed mass spectrometry, which can quickly and easily provide reference for biological and medical research.

Key words:

Quantitative proteomics
/ High resolution time-of-flight mass spectrometry
/ Counts per second
/ Signal intensity
/ Label-free quantification

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沈阳化工大学材料科学与工程学院沈阳 110142

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

通讯作者: 王勇

English

Application of High Resolution Time-of-Flight Mass Spectroscopy in Relative Quantitative Analysis in Proteomics

Corresponding author: 王勇

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通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

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

通讯作者: 王勇

English

Application of High Resolution Time-of-Flight Mass Spectroscopy in Relative Quantitative Analysis in Proteomics

Corresponding author: 王勇

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