Characterization of zebrafish cardiac proteome using online pH gradient SCX-RP HPLC-MS/MS platform.
Characterization of zebrafish cardiac proteome using online pH gradient SCX-RP HPLC-MS/MS platform.
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使用在线 pH 梯度 SCX-RP HPLC-MS/MS 平台表征斑马鱼心脏蛋白质组。
DOI：
10.1007/978-1-62703-386-2_10
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发表时间：
2013
影响因子：
--
通讯作者：
Lingjun Li
中科院分区：
文献类型：
--
作者：
Jiang Zhang;K. A. Lanham;W. Heideman;R. Peterson;Lingjun Li
二维高效液相色谱联用串联质谱（MS/MS）已成为霰弹枪蛋白质组学大规模鉴定生物样品中蛋白质的主流技术。这种强大的技术提供了速度、灵敏度和动态范围，这是从蛋白质组学样品中探测复杂肽混合物所必需的。本文提出了一种pH梯度SCX-RP 2D HPLC-MS/MS方法，旨在提高复杂蛋白质组学样品的肽分辨率和蛋白质鉴定。对pH梯度SCX-RP - 2D HPLC法与传统盐梯度SCX-RP法进行了比较。介绍了一种利用微波辅助胰蛋白酶消化的两步样品预分离方法，以提高不溶性蛋白质的鉴定。这种新的2D HPLC-MS/MS方法应用于斑马鱼的心脏蛋白质组学样本，为心血管和环境毒理学研究提供了这一重要模式生物的全面心脏蛋白质组学分析。
Two-dimensional HPLC coupled with tandem MS (MS/MS) has become a mainstream technique in the shotgun proteomics for large-scale identification of proteins from biological samples. This powerful technology provides speed, sensitivity, and dynamic range which are essential to probe complex peptide mixtures from proteomic samples. Herein we present a pH gradient SCX-RP 2D HPLC-MS/MS method designed to improve the peptide resolution and protein identification from complex proteomic samples. The comparison between the pH gradient SCX-RP 2D HPLC method and traditional salt gradient SCX-RP method was presented. A two-step sample prefractionation method utilizing microwave-assisted tryptic digestion to improve the identification of insoluble proteins was also introduced. This novel 2D HPLC-MS/MS method was applied to the heart proteomic sample of the zebrafish, Danio rerio, to provide comprehensive cardiac proteomic profiling of this important model organism for cardiovascular and environmental toxicology studies.
shot弹枪蛋白质组学的二维分级技术的比较。
影响因子：
7.4
作者：
Dowell, James A.;Frost, Dustin C.;Zhang, Jiang;Li, Lingjun
通讯作者：
Li, Lingjun