Department of Plant Nutrition, College of Resources and Environmental Sciences, China Agricultural University, Beijing, PR China.
Anal Chim Acta. 2013 Sep 17;795:82-7. doi: 10.1016/j.aca.2013.07.040. Epub 2013 Jul 26.
The tyrosine phosphorylation of proteins plays a vital role in signal transduction pathways. The highly selective enrichment of tyrosine phosphopeptides remains a significant challenge in this area of research because of the low levels of tyrosine phosphorylation in cells. Herein, we report the development of a novel molecularly imprinted polymer (MIP)-based method for the recognition of tyrosine phosphopeptides in aqueous media using an epitope approach. Phenylphosphonic acid, which has been regarded the "epitope" of phosphotyrosine, was used as a template, and commercially available zinc acrylate was used as a functional monomer to prepare the MIP. The one-pot synthetic process was simple, efficient, and the resulting MIPs were low-cost, robust, and recyclable. The MIP demonstrated significant higher levels of adsorption capacity and selectivity for phenylphosphonic acid than the non-imprinting polymer (NIP) over its structural analog benzoic acid. The MIP was also used as a molecular receptor to recognize tyrosine phosphopeptides in aqueous media, and showed a clear preference for tyrosine phosphopeptides over interfering serine peptides compared to TiO2. These results revealed the feasibility of the use of MIPs to effectively mimic the epitope approach, and provided a promising alternative to the immunoaffinity techniques commonly used for capturing tyrosine phosphopeptides.
蛋白质的酪氨酸磷酸化在信号转导途径中起着至关重要的作用。由于细胞中酪氨酸磷酸化水平较低，因此高度选择性地富集酪氨酸磷酸肽仍然是该研究领域的一个重大挑战。在此，我们报告了一种使用表位方法在水相介质中识别酪氨酸磷酸肽的新型分子印迹聚合物 (MIP) 基方法的开发。苯膦酸被认为是磷酸酪氨酸的“表位”，被用作模板，而商业可得的丙烯锌用作功能单体来制备 MIP。一锅合成过程简单、高效，所得 MIP 成本低、坚固且可回收。与结构类似物苯甲酸相比，MIP 对苯膦酸的吸附容量和选择性明显高于非印迹聚合物 (NIP)。MIP 还被用作分子受体，用于在水相中识别酪氨酸磷酸肽，与 TiO2 相比，MIP 对酪氨酸磷酸肽表现出明显的偏好，而对干扰丝氨酸肽则没有。这些结果表明，MIP 可有效地模拟表位方法，为捕获酪氨酸磷酸肽常用的免疫亲和技术提供了一种有前途的替代方法。
