Membrane protein separation and analysis by supercritical fluid chromatography-mass spectrometry.
Membrane protein separation and analysis by supercritical fluid chromatography-mass spectrometry.
复制标题
超临界流体色谱-质谱法分离和分析膜蛋白。
DOI：
10.1021/ac702319u
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发表时间：
2008
影响因子：
7.4
通讯作者：
Smith,LloydM
中科院分区：
文献类型：
--
作者：
Zhang,Xu;Scalf,Mark;Westphall,MichaelS;Smith,LloydM
膜蛋白占人类基因组的25 - 30%，在各种重要的生物过程中发挥关键作用。然而，它们的疏水性质已经损害了通过X射线晶体学和质谱法进行结构表征的努力。通常用于溶解膜蛋白的去污剂干扰X射线研究所必需的结晶过程，并导致严重的离子抑制效应，阻碍质谱分析。在这份报告中，使用超临界流体色谱-质谱的分离和分析的完整的膜蛋白和疏水肽进行了研究。结果表明，洗涤剂是快速，有效地从蛋白质和肽分离，使他们在一个适合于直接质谱分析的状态。
Membrane proteins comprise 25−30% of the human genome and play critical roles in a wide variety of important biological processes. However, their hydrophobic nature has compromised efforts at structural characterization by both X-ray crystallography and mass spectrometry. The detergents that are generally used to solubilize membrane proteins interfere with the crystallization process essential to X-ray studies and cause severe ion suppression effects that hinder mass spectrometric analysis. In this report, the use of supercritical fluid chromatography−mass spectrometry for the separation and analysis of integral membrane proteins and hydrophobic peptides is investigated. It is shown that detergents are rapidly and effectively separated from the proteins and peptides, yielding them in a state suitable for direct mass spectrometric analysis.