School of Biological Sciences and Centre for Biodiscovery, Victoria University of Wellington, Wellington, New Zealand.
Proteomics. 2011 Nov;11(22):4376-84. doi: 10.1002/pmic.201100169. Epub 2011 Oct 4.
Although there are now multiple methods for the analysis of membrane proteomes, there is relatively little systematic characterization of proteomic workflows for membrane proteins. The Asia Oceania Human Proteome Organisation (AOHUPO) has therefore embarked on a Membrane Proteomics Initiative (MPI) using a large range of workflows. Here, we describe the characterization of the MPI mouse liver microsomal membrane Standard using SDS-PAGE prior to in-gel tryptic digestion and LC-ESI-MS/MS. The Na(2) CO(3) wash followed by SDS-PAGE prior to in-gel tryptic digestion and LC-MS/MS strategy was effective for the detection of membrane proteins with 47.1% of the identified proteins being transmembrane proteins. Gene Ontology term enrichment analysis showed that biological processes involving transport, lipid metabolism, cell communication, cell adhesion, and cellular component organization were significantly enriched. Comparison of the present data with the previously published reports on mouse liver proteomes confirmed that the MPI Standard provides an excellent resource for the analysis of membrane proteins in the AOHUPO MPI.
尽管现在有多种分析膜蛋白质组的方法，但对膜蛋白质组学工作流程的系统描述相对较少。因此，亚洲大洋洲人类蛋白质组组织（AOHUPO）采用了多种工作流程，启动了膜蛋白质组学计划（MPI）。在这里，我们描述了 SDS-PAGE 之前胶内酶切和 LC-ESI-MS/MS 之前 MPI 小鼠肝微粒体膜标准的特征。Na(2)CO(3)洗涤后 SDS-PAGE 之前胶内酶切和 LC-MS/MS 策略可有效检测到 47.1%的跨膜蛋白。基因本体论术语富集分析表明，涉及运输、脂质代谢、细胞通讯、细胞黏附和细胞成分组织的生物学过程明显富集。与先前发表的关于小鼠肝蛋白质组的报告进行比较，证实 MPI 标准为 AOHUPO MPI 中膜蛋白的分析提供了极好的资源。
