College of Plant Protection, Hebei Agricultural University, Baoding 071001, PR China; State Key Laboratory of Elemento-Organic Chemistry, College of Chemistry, Nankai University, Tianjin 300071, PR China.
Hebei Key Laboratory of Plant Physiology and Molecular Pathology, Hebei Agricultural University, Baoding 071001, PR China.
Int J Biol Macromol. 2020 Dec 1;164:1475-1483. doi: 10.1016/j.ijbiomac.2020.07.237. Epub 2020 Aug 5.
Green agrochemicals are important guarantee for food production and security, and target protein identification is the most important basis for development of novel agrochemicals. Affinity chromatography methods for immobilization of agrochemicals have been widely used to identify and confirm new targets. However, this method often requires modification of the active molecules which can affect or damage its biological activity, and biomacromolecules, particularly most natural products, are hard to be modified either. In order to overcome the shortcomings of molecular modification, label-free technology has been developed based on evaluating responses to thermal or proteolytic treatments. Combined with the chemical biology technology and molecular biology technology, it has been used in the development of drugs and agrochemicals. Herein, common methods of label-free technology for identification of direct target of agrochemicals are reviewed, including the principle, advantages, limitations and applications in the research of agrochemicals in the last decade. And the methods for validation of candidate targets obtained by the label-free methods are also reviewed, which are important to obtain the accurate and reliable targets. Combined application of these methods will greatly reduce the experimental costs and shorten the period for the new target identification and validation by improving its accuracy, which will provide a systematic solution for new ecological agrochemicals research and development.
绿色农用化学品是保障粮食生产和安全的重要物质，靶标蛋白鉴定是开发新型农用化学品的重要基础。固定化农用化学品的亲和色谱法已被广泛用于鉴定和确证新靶标。然而，这种方法通常需要对活性分子进行修饰，这可能会影响或破坏其生物活性，而且生物大分子，特别是大多数天然产物，也很难进行修饰。为了克服分子修饰的缺点，基于评估对热或蛋白水解处理的响应的无标记技术已经被开发出来。该技术与化学生物学技术和分子生物学技术相结合，已被应用于药物和农用化学品的开发中。本文综述了近十年来用于鉴定农用化学品直接靶标的常见无标记技术方法，包括原理、优点、局限性及其在农用化学品研究中的应用。同时还综述了通过无标记方法获得的候选靶标的验证方法，这对于获得准确可靠的靶标非常重要。这些方法的联合应用将通过提高准确性来大大降低实验成本和缩短新靶标鉴定和验证的周期，为新型生态农用化学品的研究和开发提供系统的解决方案。
