Quantitative proteome analysis of human plasma following in vivo lipopolysaccharide administration using 16O/18O labeling and the accurate mass and time tag approach
Quantitative proteome analysis of human plasma following in vivo lipopolysaccharide administration using 16O/18O labeling and the accurate mass and time tag approach
复制标题
DOI：
10.1074/mcp.m500045-mcp200
复制
发表时间：
2005-05-01
影响因子：
7
通讯作者：
Smith, RD
中科院分区：
文献类型：
--
作者：
Qian, WJ;Monroe, ME;Smith, RD
从人血浆中鉴定新的诊断或治疗生物标志物将显著受益于在一系列生理条件下蛋白质组成分的定量测量。在这里，我们描述了一个初步的演示蛋白质组范围内的定量分析的人血浆。该方法利用消化后胰蛋白酶催化的O-16/O-18肽标记，二维LC-FTICR质谱，和准确的质量和时间（AMT）标签策略，以确定和定量肽/蛋白质复杂的样品。肽精确质量和LC洗脱时间AMT标签数据库最初使用MS/MS在广泛的多维LC分离后生成，以提供后续肽鉴定的基础。AMT标签数据库包含> 8，000个推定的已鉴定肽，提供938个可信的血浆蛋白鉴定。应用定量方法而不消耗高丰度蛋白质，用于比较分析脂多糖（LPS）给药前和给药后9 h的个体血浆样品。通过对照血浆的1：1标记和LPS给药后血浆样品之间的比较证明了蛋白质丰度变化的准确定量。从比较分析中总共定量了429种不同的血浆蛋白质，观察到25种蛋白质（包括几种已知的炎症反应介质）的蛋白质丰度在LPS给药后发生显著变化。
Identification of novel diagnostic or therapeutic biomarkers from human blood plasma would benefit significantly from quantitative measurements of the proteome constituents over a range of physiological conditions. Herein we describe an initial demonstration of proteome-wide quantitative analysis of human plasma. The approach utilizes postdigestion trypsin-catalyzed O-16/O-18 peptide labeling, two-dimensional LC-FTICR mass spectrometry, and the accurate mass and time (AMT) tag strategy to identify and quantify peptides/proteins from complex samples. A peptide accurate mass and LC elution time AMT tag data base was initially generated using MS/MS following extensive multidimensional LC separations to provide the basis for subsequent peptide identifications. The AMT tag data base contains >8,000 putative identified peptides, providing 938 confident plasma protein identifications. The quantitative approach was applied without depletion of high abundance proteins for comparative analyses of plasma samples from an individual prior to and 9 h after lipopolysaccharide (LPS) administration. Accurate quantification of changes in protein abundance was demonstrated by both 1: 1 labeling of control plasma and the comparison between the plasma samples following LPS administration. A total of 429 distinct plasma proteins were quantified from the comparative analyses, and the protein abundances for 25 proteins, including several known inflammatory response mediators, were observed to change significantly following LPS administration.