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用于磷酸化蛋白质组分析的功能化可溶性纳米聚合物

Functionalized soluble nanopolymers for phosphoproteome analysis.

作者信息

Iliuk Anton, Jayasundera Keerthi, Schluttenhofer Rachel, Tao W Andy

机构信息

Department of Biochemistry, Purdue Center for Cancer Research, Purdue University, West Lafayette, IN, USA.

出版信息

Methods Mol Biol. 2011;790:277-85. doi: 10.1007/978-1-61779-319-6_21.

Abstract

Protein phosphorylation plays a critical role in the regulation of many cellular functions. Phosphoproteomic analyses facilitate an in-depth understanding of such phosphorylation-dependent signaling networks. The use of mass spectrometry in phosphoproteomics has been especially successful, but the approach largely depends on an efficient method to enrich phosphopeptides from complex mixtures. We have developed a novel, effective soluble nanopolymer-based phosphopeptide enrichment technique, termed PolyMAC (polymer-based metal ion affinity capture). The homogenous, hyperfunctional nature of PolyMAC reagent makes it a more competent choice for highly efficient phosphopeptide binding and isolation, which was demonstrated through several applications with simple protein mixture and complex cell extract.

摘要

蛋白质磷酸化在许多细胞功能的调节中起着关键作用。磷酸化蛋白质组分析有助于深入了解此类磷酸化依赖性信号网络。质谱在磷酸化蛋白质组学中的应用尤其成功,但该方法在很大程度上依赖于从复杂混合物中富集磷酸肽的有效方法。我们开发了一种新型、有效的基于可溶性纳米聚合物的磷酸肽富集技术,称为PolyMAC(基于聚合物的金属离子亲和捕获)。PolyMAC试剂的均匀、高功能性质使其成为高效磷酸肽结合和分离的更合适选择,这在对简单蛋白质混合物和复杂细胞提取物的多次应用中得到了证明。

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