Quantitative analysis of apolipoproteins in human HDL by top-down differential mass spectrometry.
Quantitative analysis of apolipoproteins in human HDL by top-down differential mass spectrometry.
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DOI：
10.1007/978-1-62703-405-0_10
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发表时间：
2013-01-01
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影响因子：
--
通讯作者：
Cardasis, Helene L
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文献类型：
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作者：
Mazur, Matthew T;Cardasis, Helene L
随着时间的推移，定量、无标记的蛋白质组学领域已经有了显著的发展，大多数实验都是使用蛋白水解蛋白混合物“自下而上”进行的。在这些实验中，确定了两种或多种实验条件之间具有统计学意义的肽丰度差异，并随后确定其对应的蛋白质。最近，将这种实验设计扩展到完整蛋白质混合物的基本原理已经变得清晰，因为在蛋白质水平上的分析允许独立检测存在的每种蛋白质形式，包括那些翻译后修饰的蛋白质。这提供了自底向上实验所失去的特异性水平。因此，应用无标签自上而下的差分质谱法提供了一种方法来理解细微的蛋白质变化，定义一个特定的表型。这里描述的是一种方法，自上而下的无标签定量分析的蛋白质构成人高密度脂蛋白颗粒。这个方法在概念上非常简单；然而，它确实需要一定程度的严谨性和一致性，而传统的蛋白质组学实验通常无法解决这一问题。
The field of quantitative, label-free proteomics has evolved significantly over time, with most experiments performed "bottom-up" using proteolyzed protein mixtures. In these experiments, statistically significant peptide abundance differences between two or more experimental conditions are determined, and their corresponding proteins later identified. Recently, the rationale for extending this experimental design to mixtures of intact proteins has become clear, as analysis at the protein level allows for the independent detection of each protein form present, including those modified posttranslationally. This provides a level of specificity lost in bottom-up experiments. As such, the application of label-free top-down differential mass spectrometry has provided a means for understanding the subtle protein changes that define a particular phenotype. Described here is an approach for the top-down label-free quantitative analysis of the proteins which constitute human high-density lipoprotein particles. The methodology is conceptually very straightforward; however, it does require a level of rigor and consistency typically not addressed by more conventional proteomics experiments.