RAPID, EFFICIENT ELUTION OF NATIVE AFFINITY-PURIFIED PROTEIN A TAGGED COMPLEXES

本子项目是利用由NIH/NCRR资助的中心赠款提供的资源的众多研究子项目之一。子项目和研究者（PI）可能已经从另一个NIH来源获得了主要资金，因此可以在其他CRISP条目中表示。列出的机构是中心的，不一定是研究者的机构。蛋白质组学研究中面临的一个问题是如何将标记的蛋白质复合物恢复到其天然和活性形式。在这里，我们描述了一个肽，Bio-Ox，模仿免疫球蛋白G （IgG）结合界面的金黄色葡萄球菌蛋白a，并竞争性地取代亲和纯化的蛋白a融合蛋白和蛋白复合物从IgG- sepharose。我们发现Bio-Ox洗脱是一种高效快速回收天然标记蛋白的稳健方法，可应用于各种结构基因组学和蛋白质组学研究。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. A problem faced in proteomics studies is the recovery of tagged protein complexes in their native and active form. Here we describe a peptide, Bio-Ox, that mimics the immunoglobulin G (IgG) binding interface of Staphylococcus aureus Protein A, and competitively displaces affinity-purified Protein A fusion proteins and protein complexes from IgG-Sepharose. We show that Bio-Ox elution is a robust method for the efficient and rapid recovery of native tagged proteins, and can be applied to a variety of structural genomics and proteomics studies.