基于纳米结构材料的磷酸化蛋白／多肽富集和分析

Phosphoprotein/Phosphopeptide Enrichment and Analysis Based on Nanostructured Materials

蛋白质磷酸化是最为广泛的翻译后修饰之一。在生物体液或组织中,许多低丰度的磷酸化蛋白和磷酸化肽是具有高度临床灵敏性和特异性的生物标记物,这些生物分子对许多疾病的检测和病理的阐释可能提供重要的信息。因为蛋白质磷酸化动态可逆且磷酸化蛋白丰度很低,所以很难直接从复杂的生物样品中直接检测到磷酸化蛋白和磷酸化肽。纳米结构材料因其大比表面积、丰富的活性亲合位点和特殊结构,在磷酸化肽和磷酸化蛋白的分离和富集方面已经引起了特别的关注,并成为目前磷酸化蛋白质组学富集和鉴定方面的研究热点。许多介孔、磁性、杂化或化学修饰的亲合材料被研发并用于磷酸化蛋白/多肽的富集与分离;此外,一些多功能纳米结构材料也被研发并用于蛋白质组学中磷酸化蛋白/多肽的快速高效的富集提纯。在这篇综述中,我们专注于纳米结构材料在磷酸化蛋白/多肽富集和提纯方面的最新进展。

Protein phosphorytation is one of the most ubiquitous post-translational modifications. Many lowabundance endogenous phosphoproteins and phosphopeptides in the body fluids or tissues are biomarkers with higher clinical sensitivity and specificity, which could provide valuable information for the detection of many diseases and elucidation of pathology. It is still a great challenge to detect the phosphoproteins and phosphopeptides from complex biological samples directly due to reversibility of protein phosphorylation and the extremely low concentrations of phosphoproteins. Nanostructured materials have attracted particular attentions in enrichment, separation, and purification of phosphoproteins/phosphopeptides due to their larger surface area, numerous affinity sites and unique structures. The research subject has become one of research hotspots in phosphoproteomics presently. Various multifunctional nanostruetured materials such as core-shell particles with mesoporous affinity shell and magnetic core, hybrids of multiple components and composite affinity materials have been synthesized for selective enrichment and fast purification of phosphoproteins/phosphopeptides. In this review, we are focusing onrecent advancements of nanostructured materials for prior to MS analysis. Definition, structure characteris phosphoprotein/phosphopeptide enrichment and purification tic, unique physicochemical properties and potential in bioseparation of the nanostructured materials are first introduced. Subsequently, the related research background on phosphoproteomic studies in proteomics using mass spectrometric strategies in combination with phosphospecific enrichment techniques is briefly presented. After that, two types of affinity enrichment mechanisms to phosphoproteins/phosphopeptides are compactly discussed. Next, mesoporous, hybrid, and composite nanostructured materials for enrichment of phosphoproteins/phosphopeptides as well as application of muhifunctional nanostructured materials in phosphoproteomics are summarized in detail. Finally, some unsolved problems and a brief perspective and outlook on phosphopeptide enrichment are proposed.