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    • 2. 发明授权
    • Activity-based probes and methods of their preparation and use
    • 基于活性的探针及其制备和使用方法
    • US07923433B2
    • 2011-04-12
    • US10530646
    • 2003-10-08
    • David WinnDavid Alan Campbell
    • David WinnDavid Alan Campbell
    • A61K38/05
    • C07D311/90C07D311/82C12Q1/37
    • The present invention provides compositions and methods for assessing profiles of catalytically active enzymes in compositions containing a plurality of proteins. In preferred embodiments, the enzyme is a hydrolase, most preferably a cysteine protease. The methods described herein use activity based probes (“ABPs”) that have an affinity moiety for directing the binding of the ABP to one or more catalytically active target enzymes, a reactive group for forming a covalent bond at an active site of the target enzyme(s), and a TAG (e.g., a detectable label, preferably a fluorophore). One or more ABPs may be combined with a protein-containing sample under conditions for binding and reaction of the ABP(s) with target enzyme(s) that are present in the sample. The resulting products may then be used to assess the active enzyme profile of the sample, and can be correlated to the presence, amount, or activity of one or more target enzyme(s) present in the original complex protein mixture.
    • 本发明提供了用于在含有多种蛋白质的组合物中评估催化活性酶的谱的组合物和方法。 在优选的实施方案中,酶是水解酶,最优选半胱氨酸蛋白酶。 本文描述的方法使用具有亲和部分的活性基探针(“ABP”),该亲和部分用于引导ABP与一种或多种催化活性靶酶的结合,用于在靶酶的活性位点形成共价键的反应性基团 (例如,可检测标记,优选荧光团)。 可以在ABP与样品中存在的目标酶的结合和反应的条件下,将一种或多种ABP与含蛋白质的样品组合。 然后可以将所得产物用于评估样品的活性酶谱,并且可与存在于原始复合蛋白​​混合物中的一种或多种目标酶的存在,量或活性相关。