Development of receptors for N-methylated lysine- and arginine-derivatives

在这个项目中，我们建议开发超分子受体，用于基于荧光的甲基化赖氨酸和精氨酸衍生物的检测。赖氨酸和精氨酸侧链的翻译后修饰(PTM)对基因的调控和表达具有重要的作用，因此对它们的检测和定量分析工具将具有很高的价值。这些受体是专为检测部分甲基化的衍生物(如KME、Kme2、RME、sRme2)而设计的，因为目前还没有针对这些PTM的合成受体。首先，将基于BINOL-磷酸盐合成合适的初级结合基序，允许与甲基化的氨基和胍基选择性地相互作用。BINOL-磷酸允许与这些氨基酸侧链的相互作用基于几个非共价相互作用，最重要的是氢键和阳离子-pi-相互作用。然后，这些初级结合基序将被整合到多齿受体中，后者具有额外的二级结合基序。这意味着要提供足够的结合亲和力，特别是在水环境中。所得到的受体随后将用于鉴定和定量甲基化赖氨酸和精氨酸衍生物。我们将使用一种循序渐进的方法，从分离的氨基酸开始，然后是短肽，最后是整个组蛋白。如果成功，新开发的受体将首次实现对部分甲基化的赖氨酸和精氨酸的简单、基于荧光的识别，从而代表了一个有价值的(生物)分析工具。

Within this project, we suggest the development of supramolecular receptors for a fluorescence-based detection of methylated lysine- and arginine-derivatives. The posttranslational modification (PTM) of lysine and arginine sidechains has an important function for the regulation and expression of genes, so that analytical tools for their detection and quantification would be of high value. The proposed receptors are especially designed for the detection of partially methylated Derivates (such as Kme, Kme2, Rme, sRme2), because no synthetic receptors have yet been developed for these PTMs.For the development of suitable receptors, we suggest a modular concept. First, suitable primary binding motifs will be synthesized based on BINOL-phosphates, allowing a selective interaction with the methylated amino- and guanidino-groups. BINOL-phosphates allow the interaction with these amino-acid sidechains based on several noncovalent interactions, most importantly hydrogen-bonding and cation-pi-interactions. These primary binding motifs will then be integrated into multidentate receptors, which possess additional secondary binding motifs. This is meant to give sufficient binding affinities, especially in an aqueous environment. The resulting receptors will subsequently be applied for the identification and quantification of methylated lysine- and arginine-derivatives. We will use a stepwise approach, starting with isolated amino acids, followed by short peptides and finally entire histone proteins.If successful, the newly developed receptors would allow a simple, fluorescence-based identification of partially methylated lysines and arginines for the first time, thus representing a valuable (bio)analytical tool.