小鼠肝脏微粒体蛋白质组学：用于液相色谱-串联质谱分析的不同蛋白质分离工作流程的性能

Proteomics of mouse liver microsomes: performance of different protein separation workflows for LC-MS/MS.

作者信息

Zgoda Victor G, Moshkovskii Sergei A, Ponomarenko Elena A, Andreewski Timofey V, Kopylov Arthur T, Tikhonova Olga V, Melnik Stanislav A, Lisitsa Andrei V, Archakov Alexander I

机构信息

Institute of Biomedical Chemistry, Moscow, 119121, Russia.

出版信息

Proteomics. 2009 Aug;9(16):4102-5. doi: 10.1002/pmic.200900050.

DOI:10.1002/pmic.200900050

PMID:19701918

Abstract

The mouse liver microsome proteome was investigated using ion trap MS combined with three separation workflows including SDS-PAGE followed by reverse-phase LC of in-gel protein digestions (519 proteins identified); 2-D LC of protein digestion (1410 proteins); whole protein separation on mRP heat-stable column followed by 2-D LC of protein digestions from each fraction (3-D LC; 3703 proteins). The higher number of proteins identified in the workflow corresponded to the lesser percentage of run-to-run reproducibility. Gel-based method yielded a number of predicted membrane proteins similar to LC-based workflows.

摘要

使用离子阱质谱联用三种分离工作流程对小鼠肝脏微粒体蛋白质组进行了研究，这三种流程包括：SDS-PAGE 后进行凝胶内蛋白质消化的反相液相色谱法（鉴定出 519 种蛋白质）；蛋白质消化的二维液相色谱法（1410 种蛋白质）；在 mRP 热稳定柱上进行全蛋白分离，然后对每个组分的蛋白质消化产物进行二维液相色谱法（三维液相色谱法；3703 种蛋白质）。工作流程中鉴定出的蛋白质数量越多，每次运行之间的重现性百分比就越低。基于凝胶的方法产生的预测膜蛋白数量与基于液相色谱的工作流程相似。