Chemosensors for the detection of phosphate-containing biological species are in high need. Detection of proximally phosphorylated sites of PP_i and those found in peptides and proteins has been demonstrated using chemosensors containing pyrene, as a fluorescent reporter, and a Zn²⁺-chelate, as a phosphate-binding group. Using these sensors, detection of proximal phosphate groups is afforded by binding of at least two of the sensor molecules to the adjacent phosphates, via the Zn²⁺ centres, leading to excimer formation between the pyrene groups and the corresponding shift in emission from 376 to 476 nm. Although several reports of this chemosensor class have been made, no detailed studies of selectivity of these sensors among major phosphate targets have been reported. In this study, a library of this class of chemosensors, termed ProxyPhos, which contained various linkers and Zn²⁺-chelating groups (i.e. DPA, cyclen and cyclam), was prepared and the effects of structural variation on the sensing efficiency and selectivity were evaluated among proximally phosphorylated peptides, proteins, nucleotides, P_i and PP_i. As a result of this study, we have identified ProxyPhos library members that are most suitable for the detection of proximally phosphorylated peptides, PP_i, UTP, and a DpYD peptide motif, and have generally provided a foundation for the selection of ProxyPhos chemosensors for further development of specific biologically relevant assays. The broad utility of ProxyPhos is further supported by the demonstrated lack of these sensors’ cytotoxicity, ability to rapidly permeate into live and fixed cells and compatibility with gel staining methods.

中文翻译：

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用于检测近端磷酸化和其他磷酸盐种类的ProxyPhos化学传感器的结构-活性关系研究

迫切需要用于检测含磷酸盐的生物物种的化学传感器。PP的近侧磷酸化位点的检测_我和那些在肽和蛋白质中发现已经使用含有芘化学传感器证实，作为荧光报告，并且将Zn ²⁺螯合物，作为磷酸结合基团。使用这些传感器，可以通过Zn ²⁺通过将至少两个传感器分子与相邻的磷酸盐结合来提供近端磷酸盐基团的检测。中心，导致the基之间形成受激准分子，并相应地将发射从376 nm移至476 nm。尽管已经进行了有关该化学传感器类别的一些报道，但是还没有关于这些传感器在主要磷酸盐靶标中的选择性的详细研究的报道。在这项研究中，准备了一个名为ProxyPhos的此类化学传感器库，其中包含各种连接子和Zn ²⁺螯合基团（即DPA，cycln和cyclam），并且结构变化对传感效率和选择性的影响也得到了验证。在近端磷酸化的肽，蛋白质，核苷酸，P _i和PP _i中进行了评估。这项研究的结果是，我们确定了最适合检测近端磷酸化肽段，PP _i，UTP和DpYD肽基序的ProxyPhos库成员，并为进一步选择ProxyPhos化学传感器提供了基础。特定生物学相关测定的开发。ProxyPhos的广泛用途进一步得到证实，这些传感器缺乏细胞毒性，无法迅速渗透到活细胞和固定细胞中，并且与凝胶染色方法不兼容。