The development of chemical probes to study rhomboid proteases

研究菱形蛋白酶的化学探针的开发

• 批准号: 51176235
• 负责人: Professor Dr. Steven Verhelst
• 金额: --
• 依托单位: Dept. of Cellular and Molecular Medicine
• 依托单位国家: 德国
• 项目类别: Independent Junior Research Groups
• 财政年份: 2007
• 资助国家: 德国
• 起止时间: 2006-12-31 至 2013-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/51176235?language=en
• 关键词: development; chemical; probes; study; rhomboid

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.

菱形蛋白是最近发现的膜内丝氨酸蛋白酶家族，其位于脂质双层内并在其跨膜区域切割特定蛋白质。反过来，切割的蛋白质被释放并可以充当细胞间或细胞内信号。有趣的是，菱形基因存在于许多不同的生物体中，并且在整个进化过程中都是保守的。然而，仅揭示了它们的细胞作用的第一条线索。关于它们的底物特异性和在特定类型的细胞和组织中的功能作用的详细知识也缺乏。用于研究蛋白酶功能的可用化学工具与菱形不相容，可能是由于膜内蛋白水解的独特性质。在此，提出了一项计划来开发和验证选择性靶向菱形的小分子。这些分子的设计基于先前对可溶性蛋白酶小分子探针的研究，包括共价修饰剂和肽样结合元件以影响选择性。在这里，我们将丝氨酸蛋白酶反应基团与螺旋诱导元件结合起来，这些元件可以将探针引导至膜环境和菱形蛋白酶的活性位点。我们将应用这些分子来研究菱形在细菌和哺乳动物系统中的功能作用，特别是细胞通讯过程。

项目成果

Mixed alkyl aryl phosphonate esters as quenched fluorescent activity-based probes for serine proteases.

混合烷基芳基膦酸酯作为基于猝灭荧光活性的丝氨酸蛋白酶探针

• DOI: 10.1039/c4ob02444c
• 发表时间: 2015
• 期刊: Organic & biomolecular chemistry
• 影响因子: 3.2
• 作者: Verhelst;S. H. L.
• 通讯作者: S. H. L.

Activity-based probes for rhomboid proteases discovered in a mass spectrometry-based assay

• DOI: 10.1073/pnas.1215076110
• 发表时间: 2013-02-12
• 期刊: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
• 影响因子: 11.1
• 作者: Vosyka, Oliver;Vinothkumar, Kutti R.;Verhelst, Steven H. L.
• 通讯作者: Verhelst, Steven H. L.