Faculty of Pharmacy, Kindai University, 3-4-1 Kowakae, Higashi-Osaka, Osaka, 577-8502, Japan.
Graduate School of Pharmaceutical Sciences, Kyoto University, Sakyo, Kyoto, 606-8501, Japan.
Curr Top Microbiol Immunol. 2019;420:321-349. doi: 10.1007/82_2018_133.
Non-ribosomal peptide (NRP) natural products are one of the most promising resources for drug discovery and development because of their wide-ranging of therapeutic potential, and their behavior as virulence factors and signaling molecules. The NRPs are biosynthesized independently of the ribosome by enzyme assembly lines known as the non-ribosomal peptide synthetase (NRPS) machinery. Genetic, biochemical, and bioinformatics analyses have provided a detailed understanding of the mechanism of NRPS catalysis. However, proteomic techniques for natural product biosynthesis remain a developing field. New strategies are needed to investigate the proteomes of diverse producer organisms and directly analyze the endogenous NRPS machinery. Advanced platforms should verify protein expression, protein folding, and activities and also enable the profiling of the NRPS machinery in biological samples from wild-type, heterologous, and engineered bacterial systems. Here, we focus on activity-based protein profiling strategies that have been recently developed for studies aimed at visualizing and monitoring the NRPS machinery and also for rapid labeling, identification, and biochemical analysis of NRPS enzyme family members as required for proteomic chemistry in natural product sciences.
非核糖体肽 (NRP) 天然产物是药物发现和开发最有前途的资源之一，因为它们具有广泛的治疗潜力，并且作为毒力因子和信号分子发挥作用。NRP 是由酶组装线（称为非核糖体肽合成酶 (NRPS) 机器）独立于核糖体生物合成的。遗传、生化和生物信息学分析为 NRPS 催化的机制提供了详细的了解。然而，天然产物生物合成的蛋白质组学技术仍然是一个不断发展的领域。需要新的策略来研究不同产生生物体的蛋白质组，并直接分析内源性 NRPS 机器。先进的平台应验证蛋白质表达、蛋白质折叠和活性，还应能够对来自野生型、异源和工程细菌系统的生物样品中的 NRPS 机器进行分析。在这里，我们重点介绍最近为可视化和监测 NRPS 机器以及为快速标记、鉴定和生化分析 NRPS 酶家族成员而开发的基于活性的蛋白质分析策略，这些策略是天然产物科学中蛋白质组化学所必需的。
