COMPARISONS OF MASS SPECTROMETRY COMPATIBLE SURFACTANTS FOR GLOBAL ANALYSIS OF T

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 蛋白质表达的全局分析方法为研究疾病的分子基础提供了一种途径。组织如脑中蛋白质表达的研究由于需要有效和无偏的蛋白质消化而变得复杂，所述蛋白质消化允许通过鸟枪蛋白质组学方法鉴定肽。特别是，鉴定和表征丰度较低的膜蛋白对于脑生理学的研究具有极大的意义，但通常感兴趣的蛋白质丰度较低或以多种同种型存在。将蛋白质异构体作为疾病的功能进行解析将是至关重要的。在这项研究中，我们开发了一种消化方案，使用与质谱兼容的洗涤剂，提高了脑组织膜蛋白的鉴定。我们表明，修改后的程序产生接近5,000个蛋白质鉴定从1.8毫克的大鼠脑匀浆与平均25%的蛋白质序列覆盖率。该方法显著减少了观察广谱膜蛋白所需的起始材料的量。在从哺乳动物脑匀浆中鉴定的蛋白质中，1897（35%）个蛋白质被Gene Ontology注释为膜蛋白，并且1225（22.6%）个蛋白质被预测含有至少一个跨膜结构域。已鉴定的膜蛋白包括神经递质受体和与重要生理功能和疾病有关的离子通道。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Methods for the global analysis of protein expression offer an approach to study the molecular basis of disease. Studies of protein expression in tissue, such as brain, are complicated by the need for efficient and unbiased digestion of proteins that permit identification of peptides by shotgun proteomic methods. In particular, identification and characterization of less abundant membrane proteins has been of great interest for studies of brain physiology, but often proteins of interest are of low abundance or exist in multiple isoforms. Parsing protein isoforms as a function of disease will be essential. In this study, we develop a digestion scheme using detergents compatible with mass spectrometry that improves membrane protein identification from brain tissue. We show the modified procedure yields close to 5,000 protein identifications from 1.8 mg of rat brain homogenate with an average of 25% protein sequence coverage. This procedure achieves a remarkable reduction in the amount of starting material required to observe a broad spectrum of membrane proteins. Among the proteins identified from a mammalian brain homogenate, 1897 (35%) proteins are annotated by Gene Ontology as membrane proteins, and 1225 (22.6%) proteins are predicted to contain at least one transmembrane domain. Membrane proteins identified included neurotransmitter receptors and ion channels implicated in important physiological functions and disease.