生物谷報道：中科院大連化物所生物技術(shù)部1809組封順,、葉明亮,、鄒漢法等人關(guān)于新型金屬離子固定化親和色譜固定相應(yīng)用于磷酸化蛋白質(zhì)組學(xué)的研究成果(Immobilized Zirconium Ion Affinity Chromatography for Specific Enrichment of Phosphopeptides in Phosphro proteome Analysis)發(fā)表于9月份出版的《分子細胞蛋白質(zhì)組學(xué)》(Molecular & Cellular Proteomics,MCP,2007,，6，1656-1665）,?！斗肿蛹毎鞍踪|(zhì)組學(xué)》由美國生物化學(xué)和細胞生物學(xué)學(xué)會主辦，是蛋白質(zhì)組學(xué)研究領(lǐng)域最高影響因子的學(xué)術(shù)刊物,，2005年和2006年的SCI引用影響因子分別為9.876和9.62,。
在復(fù)雜的蛋白質(zhì)提取物的酶解產(chǎn)物中，磷酸化多肽的高選擇性,、高穩(wěn)定性富集是磷酸化蛋白質(zhì)組學(xué)分析的關(guān)鍵步驟之一,。常規(guī)的金屬離子固定化親和色譜固定相以氨基二乙酸(IDA)或次氮基三乙酸(NTA)為螯合基團與Fe3+,，Ga3+等離子螯合,，進而選擇性地作用于磷酸化肽段,，達到富集磷酸化多肽的目的。這一類吸附材料對含有酸性氨基酸殘基側(cè)鏈的肽段也具有富集作用,，因此嚴重干擾磷酸化肽段的檢測,。本項工作制備了磷酸脂改性的高分子材料微球，利用磷酸脂基團與鎬離子的配位作用固定化鎬離子,，再通過鎬離子與磷酸肽上的磷酸基團的選擇性作用達到富集的目標(biāo),。實驗結(jié)果表明鎬離子固定化親和色譜固定相對磷酸化肽段具有很高選擇性和富集效果。（援引中科院大連化物所）
原始出處:
Molecular & Cellular Proteomics 6:1656-1665, 2007.
Immobilized Zirconium Ion Affinity Chromatography for Specific Enrichment of Phosphopeptides in Phosphoproteome Analysis*,S
Shun Feng,,¶, Mingliang Ye,¶, Houjiang Zhou, Xiaogang Jiang, Xingning Jiang, Hanfa Zou,|| and Bolin Gong**
From the National Chromatographic Research and Analysis Center, Dalian Institute of Chemical Physics, The Chinese Academy of Sciences, Dalian 116023, China, College of Chemistry and Chemical Engineering, Xinjiang University, Urumqi, Xinjiang 830046, China, and ** Key Laboratory of Biotechnology, Ningxia University, Yin chuan 750021, China
Large scale characterization of phosphoproteins requires highly specific methods for purification of phosphopeptides because of the low abundance of phosphoproteins and substoichiometry of phosphorylation. Enrichment of phosphopeptides from complex peptide mixtures by IMAC is a popular way to perform phosphoproteome analysis. However, conventional IMAC adsorbents with iminodiacetic acid as the chelating group to immobilize Fe3+ lack enough specificity for efficient phosphoproteome analysis. Here we report a novel IMAC adsorbent through Zr4+ chelation to the phosphonate-modified poly(glycidyl methacrylate-co-ethylene dimethacrylate) polymer beads. The high specificity of Zr4+-IMAC adsorbent was demonstrated by effectively enriching phosphopeptides from the digest mixture of phosphoprotein (- or ß-casein) and bovine serum albumin with molar ratio at 1:100. Zr4+-IMAC adsorbent was also successfully applied for the analysis of mouse liver phosphoproteome, resulting in the identification of 153 phosphopeptides (163 phosphorylation sites) from 133 proteins in mouse liver lysate. Significantly more phosphopeptides were identified than by the conventional Fe3+-IMAC approach, indicating the excellent performance of the Zr4+-IMAC approach. The high specificity of Zr4+-IMAC adsorbent was found to mainly result from the strong interaction between chelating Zr4+ and phosphate group on phosphopeptides. Enrichment of phosphopeptides by Zr4+-IMAC provides a powerful approach for large scale phosphoproteome analysis.
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