Chemical Biology Approaches Reveal Conserved Features of a C‐Terminal Processing PDZ Protease
Chemical Biology Approaches Reveal Conserved Features of a C‐Terminal Processing PDZ Protease
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DOI：
10.1002/cbic.201100643
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发表时间：
2012-02
期刊：
影响因子：
3.2
通讯作者：
Juliane Weski;M. Meltzer;Lina Spaan;T. Mönig;Julian Oeljeklaus;P. Hauske;Lars Vouillème;R. Volkmer;P. Boisguérin;D. Boyd;R. Huber;M. Kaiser;M. Ehrmann
中科院分区：
文献类型：
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作者：
Juliane Weski;M. Meltzer;Lina Spaan;T. Mönig;Julian Oeljeklaus;P. Hauske;Lars Vouillème;R. Volkmer;P. Boisguérin;D. Boyd;R. Huber;M. Kaiser;M. Ehrmann
高温需要量A （HtrA）蛋白家族含有内部或C端PDZ结构域，在革兰氏阴性菌包膜蛋白质量控制中起关键作用。虽然几种HtrA蛋白酶已被广泛表征，但C末端加工蛋白酶的许多特征，如尾部特异性蛋白酶（Tsp）仍然未知。为了充分了解这些细胞控制系统，个体结构域需要被特定的肽作为激活剂或抑制剂。在这里，我们描述了有效的抑制剂和Tsp激活剂的鉴定和设计。确定了合适的Tsp合成底物，并为生成基于硼酸的肽抑制剂提供了基础。此外，利用合成肽库对大肠杆菌细胞包膜蛋白进行蛋白质组学筛选，以鉴定能够扩增Tsp蛋白水解活性的肽。讨论了这些发现对PDZ蛋白酶调控和未来机制研究的意义。
Several proteases like the high temperature requirement A (HtrA) protein family containing internal or C‐terminal PDZ domains play key roles in protein quality control in the cell envelope of Gram‐negative bacteria. While several HtrA proteases have been extensively characterized, many features of C‐terminal processing proteases such as tail‐specific protease (Tsp) are still unknown. To fully understand these cellular control systems, individual domains need to be targeted by specific peptides acting as activators or inhibitors. Here, we describe the identification and design of potent inhibitors and activators of Tsp. Suitable synthetic substrates of Tsp were identified and served as a basis for the generation of boronic acid‐based peptide inhibitors. In addition, a proteomic screen of E. coli cell envelope proteins using a synthetic peptide library was performed to identify peptides capable of amplifying Tsp's proteolytic activity. The implications of these findings for the regulation of PDZ proteases and for future mechanistic studies are discussed.