A Conjugated Polymer Fluorogenic Probe for Inorganic Polyphosphate

描述（由申请人提供）：聚磷酸盐（poly P）是一种无机磷酸盐的线性聚合物，存在于所有生物体中，长度从数十到数百个残基不等。虽然其生物学功能尚未完全阐明，但最近的工作已经阐明了其在病原体毒力，血凝级联，生物矿化和金属隔离中的作用。正在进行的研究表明，多聚P长度与其起源、功能和酶促环境有关。然而，现有的聚P分析是简单的，需要最小的样品纯化不能提供长度的信息。长度特定的方法不适合高通量格式，需要大量的纯化或耗时的步骤。为了加快基础生化研究和发现靶向多聚磷介导途径的药物，需要简便、敏感和长度特异性的测定方法。本方案描述了一种长度特异性的、基于共轭聚合物的聚磷荧光探针。在没有聚磷的情况下，共轭聚合物被淬灭。聚P结合到一个柔性段诱导构象变化，恢复荧光。poly - P结合区域的长度可以定制，使长度选择性荧光测定的发展成为可能。分子识别机制的灵感来自于一个结构独特的离子通道，在许多细菌中发现，由聚酯、Ca2+和聚p组成。使用非离子探针将最大限度地减少与其他生物分子的非特异性相互作用。对聚磷和荧光探针之间相互作用的详细研究将为优化分析提供信息，并为分子识别提供见解。所述探针的预期选择性将使其能够用于自然样品的分析。除了促进聚磷生物化学的研究外，这些研究将为聚磷生物化学的研究提供一个新的途径

DESCRIPTION (provided by applicant): Polyphosphate (poly P) is a linear polymer of inorganic phosphates that is found in all living organisms, ranging from tens to hundreds of residues in length. Although its biological functions have not been fully elucidated, recent work has shed light on its role in the virulence of pathogens, the blood clotting cascade, biomineralization, and metal sequestration. Ongoing research suggests that poly P length is correlated with its origin, function, and enzymatic environment. However, existing poly P assays that are facile and require minimal sample purification do not provide information about length. Methods that are length specific are not suitable to a high-throughput format, requiring extensive purification or time-consuming steps. In order to expedite fundamental biochemical studies and the discovery of drugs that target poly P-mediated pathways, facile, sensitive, and length-specific assays are needed. This proposal describes a length-specific, conjugated polymer-based fluorogenic probe for poly P. In the absence of poly P, the conjugated polymer is quenched. Poly P binding to a flexible segment induces a conformational change that restores fluorescence. The length of the poly P- binding region may be tailored, enabling the development of a length-selective fluorescence assay. The mechanism of molecular recognition is inspired by a structurally unique ion channel, found in many bacteria, composed of a polyester, Ca2+, and poly P. The use of a nonionic probe will minimize nonspecific interactions with other biomolecules. Detailed studies of the interaction between poly P and the fluorogenic probe will inform optimization of the assay and provide insights into molecular recognition. The anticipated selectivity of the described probe will enable its use for the analysis of natural samples. In addition to facilitating the study of poly P biochemistry, these studies will provide a framework for the development of chemosensors for other polyelectrolytes.