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

快速、高效地洗脱天然亲和纯化的蛋白 A 标记复合物

• 批准号: 7355138
• 负责人: MICHAEL P ROUT
• 金额: $ 1.23万
• 依托单位: ROCKEFELLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-03-01 至 2007-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7355138
• 关键词: RAPID; EFFICIENT; ELUTION; NATIVE; AFFINITY

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.

该子项目是利用NIH/NCRR资助的中心赠款提供的资源的许多研究子项目之一。子项目和研究者（PI）可能从另一个NIH来源获得主要资金，因此可以在其他CRISP条目中表示。所列机构为中心，不一定是研究者所在机构。蛋白质组学研究中面临的一个问题是以其天然和活性形式回收标记的蛋白质复合物。在这里，我们描述了一种肽，Bio-Ox，它模拟金黄色葡萄球菌蛋白A的免疫球蛋白G（IgG）结合界面，并竞争性地取代亲和纯化的蛋白A融合蛋白和IgG-琼脂糖凝胶的蛋白复合物。我们表明，Bio-Ox洗脱是一种有效和快速回收天然标记蛋白质的稳健方法，可应用于各种结构基因组学和蛋白质组学研究。