Selective activity-based probes for cysteine cathepsins
Selective activity-based probes for cysteine cathepsins
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基于活性的半胱氨酸胰蛋白酶选择性探针
DOI：
10.1002/anie.200702811
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发表时间：
2008-01-01
影响因子：
16.6
通讯作者：
Wendt, K. Ulrich
中科院分区：
文献类型：
--
作者：
Watzke, Anja;Kosec, Gregor;Wendt, K. Ulrich
蛋白水解酶在许多重要的过程中起着至关重要的作用，对细胞信号和组织动态平衡是至关重要的。[1-3]由于蛋白水解酶的活性必须受到严格的控制，所以大多数的蛋白水解酶都是以不活跃的前体(酵素)的形式合成的，通过有限的蛋白水解酶来激活。因此，大多数经典的生物化学和蛋白质组学方法不能区分特定的蛋白水解酶在体内的表达水平和活性。因此，近几年来，利用化学手段对蛋白质酶活性而不是对其表达进行生物分子成像已经引起了人们的极大兴趣，并开发了一系列基于活性的内猝灭探针(ABPs)。[4，5]这些探针在蛋白水解酶的裂解后变成荧光。要分析一种独特的蛋白酶在细胞复杂环境中的功能，需要开发选择性的和细胞通透性的ABPs，这一直是具有挑战性的。目前，用于蛋白水解酶的内部猝灭的成像探针已经从多肽底物中衍生出来，耦合到适当的报告基团，这些报告基团在同源序列的蛋白分解裂解时在空间上分离。[4]优选的底物通常从位置扫描组合文库中识别。[6]这样的文库的合成和扫描可能是耗时的，并且当探针应用于细胞或整个动物成像实验时，所得到的底物的多肽性质可能导致其他蛋白酶的非特异性切割。对于半胱氨酸组织蛋白家族的蛋白酶，基于细胞渗透活性的探针被设计为自杀亚
Proteases are critically involved in a multitude of vital processes, essential for cellular signaling and tissue homeostasis.[1–3] Since proteolytic activity has to remain under stringent control, most of the proteases are synthesized as inactive precursors (zymogens), which are activated by limited proteolysis. Consequently, most of the classical biochemical and proteomics methods fail to discriminate between the expression level and proteolytic activity of a given protease in vivo. Therefore, the development of chemical tools for biomolecular imaging of protease activity rather than protease expression has gained considerable interest in recent years, and a series of internally quenched activity-based probes (ABPs) for proteases has been developed.[4, 5] These probes become fluorescent upon proteolytic cleavage.Analysis of the function of a distinct protease in the complex milieu of the cell requires the development of selective and cell-permeable ABPs, which has been challenging. To present, internally quenched imaging probes for proteolytic enzymes have been derived from peptidic substrates, coupled to appropriate reporter groups which are spatially separated upon proteolytic cleavage of the cognate sequence.[4] The preferred substrates are commonly identified from positional scanning combinatorial libraries.[6] The synthesis and scanning of such libraries can be time consuming, and the peptidic nature of the resulting substrates may result in unspecific cleavage by other proteases when the probes are applied in cellular or whole animal imaging experiments. For proteases from the cysteine cathepsin family, cell-permeable activity-based probes have been designed as suicide sub-