Subtle Modification of Isotope Ratio Proteomics (SMIRP)

同位素比蛋白质组学的微妙修改 (SMIRP)

基本信息

批准号：
7770893
负责人：
Julian P Whitelegge
金额：
$ 19.06万
依托单位：
UNIVERSITY OF CALIFORNIA LOS ANGELES
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-03-22 至 2012-02-28
项目状态：
已结题

项目摘要

DESCRIPTION (provided by applicant): Proteomics seeks to monitor the flux of protein through a biological system under variable developmental and environmental influences as programmed by the genome but, if it is to deliver its true potential, it is critically important that our proteomics technologies deliver reproducible quantitation. Because of the variability of day-to-day measurements of protein quantities, a common feature of quantitative proteomics is the use of stable isotope coding to distinguish control and experimental samples in a mixture that can be profiled in a single experiment. Coding with stable isotopes can be achieved by growth of an organism in depleted/enriched media, or by chemically modifying proteins after extraction from the organism, though these later approaches are not useful for measurement of protein turnover rates. Current stable isotope strategies seek full isotope- exchange, that is, to swap all 14N for 15N, 12C for 13C, or 16O for 18O, such that a peptide's mass is altered by several Daltons. These methods are expensive, because of the need for high isotope purity, and it was estimated that the first 15N rat cost ~$10,000. Furthermore, the fact that two peptide isotopomer distributions replace one leads to a practical loss of separation space in the mass spectrometer demanding more efficient peptide separations. Moreover, the second isotopomer distribution may trigger MS-MS in automated proteomics experiments, wasting mass spectrometer time. If a particular protein has been massively up-or-down-regulated there is also the possibility that its presence will be ignored because there weren't any readily identified pairs of peptide signals in the mass spectrum. We propose here an inexpensive strategy, potentially applicable to expression proteomics and turnover measurements in living humans, that codes the quantitative differential expression information within a single isotopomer envelope.

描述（由申请人提供）：蛋白质组学试图通过基因组编程的可变发育和环境影响来监测蛋白质通过生物系统的通量，但如果要发挥其真正的潜力，那么我们的蛋白质组学技术至关重要，我们的蛋白质组学技术提供了可重复的定量。由于蛋白质量的日常测量值的可变性，定量蛋白质组学的一个共同特征是使用稳定的同位素编码来区分混合物中可以在单个实验中介绍的混合物中的控制和实验样品。用稳定的同位素编码可以通过在耗尽/富集的培养基中的生长或从生物体提取后的化学修饰蛋白质来实现，尽管这些后来的方法对于测量蛋白质周转率并不有用。当前的稳定同位素策略寻求完全同位素交换，也就是说，将所有14N换成15N，为13c或16o将所有同位素交换为18O，以便将肽的质量改变了几个daltons。由于需要高同位素纯度，这些方法很昂贵，据估计，第一个15N老鼠的成本约为10,000美元。此外，两个肽同位素分布取代了一个事实导致质谱仪中分离空间的实际损失，要求更有效的肽分离。此外，第二个同位素分布可能触发自动蛋白质组学实验中的MS-MS，浪费质谱仪时间。如果特定的蛋白质被大规模上调节或下调，则可能会忽略它的存在，因为在质谱中没有任何容易鉴定出的成对的肽信号。我们在这里提出了一种廉价的策略，该策略可能适用于活着的人类的表达蛋白质组学和周转率测量，该策略代码单个同位素包膜内的定量差分表达信息。

项目成果

期刊论文数量（6）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

The cell wall and secretory proteome of a tobacco cell line synthesising secondary wall.

合成次生壁的烟草细胞系的细胞壁和分泌蛋白质组。

DOI：
10.1002/pmic.200800721
发表时间：
2009
期刊：
Proteomics
影响因子：
3.4
作者：
Millar,DavidJ;Whitelegge,JulianP;Bindschedler,LaurenceV;Rayon,Catherine;Boudet,Alain-Michel;Rossignol,Michel;Borderies,Gisèle;Bolwell,GPaul
通讯作者：
Bolwell,GPaul

Diverse facets of plant proteomics.

植物蛋白质组学的各个方面。