A Conjugated Polymer Fluorogenic Probe for Inorganic Polyphosphate

无机聚磷酸盐的共轭聚合物荧光探针

• 批准号: 8662083
• 负责人: Julia Ann Kalow
• 金额: $ 5.15万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-08-01 至 2016-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8662083
• 关键词: Bacteria; Binding; Biochemical; Biochemistry; Biogenesis; Biological Assay; Biological Process; Blood coagulation; Chemicals; Complex Mixtures; Detection; Development; Drug Targeting; Environment; Fluorescence; Fluorescent Probes; Goals; Ion Channel; Ions; Length; Life; Light; Mediating; Metals; Methods; Organism; Osteogenesis; Pathway interactions; Play; Polyesters; Polymers; Polyphosphates; Research; Research Personnel; Role; Sampling; Sensory; Specificity; Structure; Time; Virulence; Work; base; biomineralization; copolymer; design; flexibility; inorganic phosphate; insight; molecular recognition; novel therapeutics; pathogen; public health relevance

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.

描述（由申请人提供）：聚磷酸盐（poly P）是无机磷酸盐的线性聚合物，存在于所有活生物体中，长度范围为数十至数百个残基。虽然其生物学功能尚未完全阐明，但最近的工作揭示了其在病原体毒力，凝血级联，生物矿化和金属螯合中的作用。正在进行的研究表明，聚P的长度与其来源，功能和酶环境相关。然而，现有的多聚P测定是容易的，需要最少的样品纯化不提供有关长度的信息。长度特异性的方法不适合高通量形式，需要大量纯化或耗时的步骤。为了加快基础生物化学研究和靶向聚P介导的途径的药物的发现，需要简便、灵敏和长度特异性的测定。该提案描述了一种针对聚P的长度特异性、基于共轭聚合物的荧光探针。在不存在聚P的情况下，共轭聚合物被猝灭。聚P与柔性片段的结合诱导恢复荧光的构象变化。聚P-结合区的长度可以定制，使得能够开发长度选择性荧光测定。分子识别的机制受到结构独特的离子通道的启发，在许多细菌中发现，由聚酯，Ca 2+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。聚P和荧光探针之间的相互作用的详细研究将通知优化的测定，并提供对分子识别的见解。所述探针的预期选择性将使其能够用于天然样品的分析。除了促进聚磷的生物化学研究，这些研究将提供一个 其他聚电解质的化学传感器的发展框架。