Rapid Affinity Enrichment of Human Apolipoprotein A-I Associated Lipoproteins for Proteome Analysis,Journal of Proteome Research

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Rapid Affinity Enrichment of Human Apolipoprotein A-I Associated Lipoproteins for Proteome Analysis
Journal of Proteome Research ( IF 3.8 ) Pub Date : 2018-02-13 00:00:00 , DOI: 10.1021/acs.jproteome.7b00816
Timothy S. Collier ₁ , Zhicheng Jin ₁ , Celalettin Topbas ₁ , Cory Bystrom ₁

Affiliation

Isolation of high density lipoproteins (HDL) for structural and functional studies typically relies on ultracentrifugation techniques, which are time-consuming and difficult to scale. With emerging interest in the clinical relevance of HDL structure and function to cardiovascular disease, a significant gap exists between current and desirable sample preparation throughput. To enable proteomic studies of HDL with large clinical cohorts, we have developed an affinity enrichment approach that relies on the association of histidine-tagged, lipid free ApoA-I with HDL followed by standard metal chelate chromatography. Characterization of the resulting affinity-enriched ApoA-I associated lipoprotein (AALP) pool using biochemical, electrophoretic, and proteomic analysis demonstrates that the isolated material is closely related in structural features, lipid content, protein complement, and relative protein distribution to HDL isolated by ultracentrifugation using sequential density adjustment. The simplicity of the method provides avenues for high-throughput analysis of HDL associated proteins.

中文翻译：

人类载脂蛋白AI相关脂蛋白的快速亲和富集用于蛋白质组分析

用于结构和功能研究的高密度脂蛋白（HDL）的分离通常依赖于超速离心技术，该技术费时且难以扩展。随着人们对HDL结构和功能与心血管疾病的临床相关性产生了新的兴趣，当前和理想的样品制备通量之间存在巨大差距。为了能够对具有大量临床队列的HDL进行蛋白质组学研究，我们开发了一种亲和力富集方法，该方法依赖于组氨酸标签的，无脂质的ApoA-I与HDL的结合，然后进行标准的金属螯合层析。使用生化，电泳和蛋白质组学分析对所得的亲和力丰富的ApoA-I相关脂蛋白（AALP）库进行表征，表明分离出的物质在结构特征上密切相关，脂质含量，蛋白质补体和相对蛋白质相对于HDL的分布（通过使用顺序密度调节进行超速离心分离）。该方法的简便性为HDL相关蛋白的高通量分析提供了途径。

更新日期：2018-02-14

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