Chemical proteomic strategies for deciphering neocarzilin´s mode of action in cancer cells

破译新卡齐林在癌细胞中作用模式的化学蛋白质组学策略

• 批准号: 426512676
• 负责人: Dr. Sabine Schneider
• 金额: --
• 依托单位: Department Chemie
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2019
• 资助国家: 德国
• 起止时间: 2018-12-31 至 2023-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/426512676?language=en
• 关键词: Chemical; proteomic; strategies; deciphering; neocarzilin

Natural products address a plethora of different proteins with essential functions. The deconvolution of their targets represents a unique opportunity for unravelling new anti-cancer strategies. We here apply chemical proteomics to identify the targets of the natural product neocarzilin within living cancer cells. Neocarzilin is a potent anti-cancer compound which was identified decades ago but still lacks functional characterization. In preliminary studies we synthetically equipped the natural product with an alkyne tag and treated proteomes to identify its cellular target(s). Interestingly, only one protein, the vesicle amine transport protein (VAT-1) with an essential role in cell migration, has been identified as prominent hit. As neocarzilin also exhibits strong anti-proliferative effects which are independent of VAT-1, other yet unidentified targets must exist. To decipher these targets we aim to improve our initial probe by incorporation of a signature stereocenter present in the natural product but absent in the first generation probe design. Target identification will follow our established chemical proteomic procedures and selected protein hits will be validated for compound binding by overexpression and crystallography. Another strong focus of this work is the already identified hit VAT-1. Our initial validation revealed a putative role in cancer cell migration, an important feature of malignant tumors. Further chemical proteomic studies are required to understand the cellular function of this poorly characterized protein and to exploit its potential for future cancer treatment. The studies proposed here will include co-immunoprecipitation in presence and absence of the natural product to decipher interaction partners, co-crystallization and structure elucidation with the compound to unravel the binding mode and rationalize improved binders. The overall aims of this proposal are to decipher the full spectrum of neocarzilin targets, understand its mode of action, investigate the role of VAT-1 and find improved derivatives with therapeutic potential.

天然产品介绍了具有重要功能的大量不同蛋白质。其目标的反卷积代表了揭示新的反癌策略的独特机会。我们在这里使用化学蛋白质组学来识别活癌细胞中天然产物新核酸盐的靶标。 Neocarzilin是一种有效的抗癌化合物，几十年前被发现，但仍然缺乏功能表征。在初步研究中，我们在合成的天然产物中配备了炔烃标签和处理过的蛋白质组织，以鉴定其细胞靶标。有趣的是，只有一种蛋白质，即在细胞迁移中起着至关重要的作用的囊泡胺转运蛋白（VAT-1）。由于新可甲酶也表现出强烈的抗增殖作用，而这些作用与VAT-1无关，因此必须存在其他未识别的靶标。为了破译这些目标，我们旨在通过合并天然产品中存在的签名立体中心，但在第一代探针设计中不存在。靶标识别将遵循我们已建立的化学蛋白质组学程序，并且通过过表达和晶体学的化合物结合进行了验证。这项工作的另一个重点是已经确定的HIT VAT-1。我们的最初验证表明，在癌细胞迁移中起着推定的作用，这是恶性肿瘤的重要特征。需要进一步的化学蛋白质组学研究来了解这种表征不佳的蛋白质的细胞功能，并利用其未来癌症治疗的潜力。此处提出的研究将包括在存在和不存在天然产物的情况下进行免疫沉淀，以破译相互作用伙伴，共结晶和结构阐明与化合物阐明结合模式并合理化改进的粘合剂。该提案的总体目的是破译新的Neocarzilin靶标，了解其作用方式，研究VAT-1的作用，并发现具有治疗潜力的改进衍生物。