a Laboratory of Drug Discovery and Molecular Engineering, Department of Medicinal Plants, College of Plant Science and Technology , Huazhong Agricultural University (HZAU) , Wuhan , China.
b National & Local Joint Engineering Research Center (Hubei) for Medicinal Plant Breeding and Cultivation , Wuhan , China.
Artif Cells Nanomed Biotechnol. 2018;46(sup2):386-396. doi: 10.1080/21691401.2018.1457539. Epub 2018 Apr 3.
With the advantages of unicellular eukaryotic structure and easy manipulation, yeast becomes a popular tool for biochemical, genetic and medicinal studies. In order to construct an efficient anti-inflammatory drug screening platform, we engineered yeast as a double-molecule carrier, of which an inserted domain (I domain) of lymphocyte function-associated antigen 1 was displayed on yeast surface and a green fluorescent protein (GFP) was expressed inside cytosol. The I domain specifically targeted a surface marker of mammalian cells, intercellular adhesion molecule 1, whose number is correlated with the level of cellular inflammation. Examination of GFP intensity enables swift quantification of the yeast-mammalian cell binding and thus it reflects inflammatory potency, herein the inflammatory index, of a chemical imposed to cells. The inflammatory potency of a total of 1340 chemicals was indexed. Among them, 1 inflammation-inducing and 1 inflammation-reducing compounds were verified both in vitro and in vivo. Our method demonstrated a swift, facile and high-throughput screening platform at the protein level for inflammation and related diseases drug discovery without using sophisticated instruments.
酵母具有单细胞真核生物结构的优势，且易于操作，因此成为生化、遗传和医学研究的热门工具。为了构建高效的抗炎药物筛选平台，我们将酵母设计为双分子载体，其中淋巴细胞功能相关抗原 1 的插入结构域（I 结构域）展示在酵母表面，绿色荧光蛋白（GFP）在细胞质内表达。I 结构域特异性靶向哺乳动物细胞的表面标志物细胞间黏附分子 1，其数量与细胞炎症水平相关。通过检测 GFP 强度，可以快速定量酵母-哺乳动物细胞的结合，从而反映化学物质对细胞的炎症活性，即炎症指数。我们对总共 1340 种化学物质的炎症活性进行了指数评估。其中，1 种诱导炎症的化合物和 1 种抗炎化合物在体外和体内都得到了验证。我们的方法无需使用复杂的仪器，在蛋白质水平上展示了一种快速、简便、高通量的炎症及相关疾病药物筛选平台。
