The development of chemical probes to study rhomboid proteases

菱形蛋白酶是近年来发现的一类膜内丝氨酸蛋白酶，它位于脂质双分子层内，切割跨膜区的特异性蛋白。反过来，叶子蛋白被释放出来，并作为细胞内或细胞间的信号。有趣的是，菱形基因存在于许多不同的生物体中，并在整个进化过程中被保留下来。然而，关于它们在细胞中的作用，目前只发现了第一条线索。关于它们在特定类型的细胞和组织中的底物特异性和功能作用的详细知识也缺乏。用于研究蛋白酶功能的现有化学工具与菱形蛋白不相容，这可能是由于膜内蛋白水解的排他性。在这里，提出了一个计划，以开发和验证小分子，选择性目标菱形。这些分子的设计是基于以前的工作的小分子探针的可溶性蛋白酶，包括共价修饰剂和肽类binding元素，以影响选择性。在这里，我们将联合收割机丝氨酸蛋白酶反应基团与螺旋诱导元件相结合，该螺旋诱导元件可以将探针引导到膜环境和菱形蛋白酶的活性位点。我们将把这些分子应用于研究菱形在细菌和哺乳动物系统中的功能作用，特别是关注细胞通讯过程。

Rhomboids are a recently discovered family of intra-membrane serine proteases that residewithin a lipid bilayer and cleave specific proteins in their trans-membrane region. In turn, thecleaved protein is released and can act as an inter- or intracellular signal.Interestingly, rhomboid genes are found in many diverse organisms and are conservedthroughout evolution. However, only the first clues to their cellular roles have been uncovered.Detailed knowledge about their substrate specificity and functional roles in specific types ofcells and tissues is also lacking.The available chemical tools that have been used to study protease function areincompatible with rhomboids, possibly due to the exclusive nature of intra-membraneproteolysis. Here, a plan is proposed to develop and validate small molecules that selectivelytarget rhomboids. The design of these molecules is based on previous work on smallmolecule probes for soluble proteases, comprising of a covalent modifier and a peptide-likebinding element to influence selectivity. Here, we will combine serine protease reactivegroups with helical-inducing elements that can direct the probes to the membraneenvironment and the active site of the rhomboid protease. We will apply these molecules inthe study of the functional role of rhomboid in bacterial and mammalian systems, particularlyfocusing on cell communication processes.