Fluorogenic imaging molecules for protease/kinase assays

描述(由申请人提供)：这项建议的目的是开发一种 一系列荧光成像分子，可以很容易地输送到 活细胞和监测细胞功能。电影是由一种多肽组成的。 含有一种激酶或一种蛋白酶的靶点的骨架。一些设计 借用分子信标方法携带荧光团/猝灭剂 部分，而其他设计只携带一个荧光团。当出现时， 淬火部分是FIM内部的，而不是在极端的N-AND C-端，与其他类似类型的信标的情况一样。这避免了 结构约束的必要性，其目的是为了带来 从而使荧光团和猝灭剂彼此紧密接近 防止背景荧光。具体地说，这项提案的范围 将介绍两种多肽的设计、合成和测试 Caspase-1(ICE)和caspase-3(CPP32)的切割位点。也是 这项提案包括设计、合成和测试含氟化合物 来源于差异磷酸化的多肽，必需的结构域 人cdd25C磷酸酶，参与有丝分裂的诱导 进程。

DESCRIPTION (provided by applicant): The aim of this proposal is to develop a series of fluorogenic imaging molecules, which can be easily delivered into viable cells and monitor cellular functions. The FIMs are composed of a peptide backbone containing a target site for a kinase or a protease. Some designs borrow from the molecular beacon approach and carry flurophore/quencher moieties, while other designs simply carry a flurophore only. When present, the quencher moiety is internal to the FIM rather than at the extreme N- and C-terminus, as has been the case with other similar type beacons. This avoids the need for structural constraint, which aims to be imposed in order to bring the flurophore and quencher into close proximity to each other thereby preventing background fluorescence. Specifically, the scope of this proposal will cover the design, synthesis and testing of two peptides containing cleavage sites specific for caspase-1 (ICE) and caspase-3 (CPP32). Also included in this proposal is the design, synthesis and testing of fluorogenic peptides derived from a differentially phosphorylated, essential domain of human cdd25C phosphatase, which is involved in induction of the mitotic process.