The new method to identify small molecules binding proteins by usingmutants and genome sequences

本研究旨在鉴定新的作用蛋白和体内靶蛋白，建立利用基因突变的黑腹果蝇和使用它的CPT的新药接受性评价方法。CPT-20-B连接肽（NSASRGGSQRGRGEH）采用晶体偏离摆微天平（QCM）装置T7噬菌体展示法进行鉴定。该肽序列显示了人类HshnRNP A1和果蝇DmHrb87F的同源性。CPT联合从头部下拉检查，使用海拉细胞熔融图像与HshnRNP A1。CPT制备HshnRNP A1- gst联合变化蛋白，进行QCM互作分析，为HshnRNP A1，解离常数为82.7 nM。微生物敏感性试验结果表明，通过基因丢失，两个DmHrb87F基因突变体Hrb87FKG02089和Hrb87FBG02743与野生菌株相比表现出CPT高接受度。Topo I的工作是消除质粒DNA的超级线圈。这种拓扑变化是在CPT和HshnRNP A1存在的情况下观察到的。正如报道的那样，CPT阻碍了topo I的振兴。此外，hnRNP A1也同样阻碍了topo I的振兴。此外，两种情况下，CPT均改善了topo I对HshnRNP A1的活性抑制。HshnRNP A1或DmHrb87F与CPT结果一致，并提出了调整topo I活化的可能性。

The present study aimed at the identification of the action new and in vivo target protein of the establishment of the new medicine receptivity evaluation method that used gene mutation Drosophila melanogaster and CPT that used it.CPT-20-B connecting peptide (NSASRGGSQRGRGEH) is identified by using the T7 phage display method with crystal departure pendulum microbalance (QCM) device. This peptide sequencing showed each array and the homology of human HshnRNP A1 and Drosophila melanogaster DmHrb87F. CPT united from the bead pull-down examination that used HeLa cell meltable picture with HshnRNP A1. CPT made HshnRNP A1-GST united changing protein, did the QCM interaction analysis, and was HshnRNP A1 and was dissociation constant 82.7 nM.As a result of the microbial sensitivity test, by gene loss Drosophila melanogaster, two Hrb87FKG02089 and Hrb87FBG02743 that was the DmHrb87F gene mutant showed CPT high receptivity compared with the wild strain. Topo I has working that cancels a super-coil of plasmid DNA. This topology change was observed under CPT and the HshnRNP A1 existence. CPT obstructed the topo I revitalization as reported. Moreover, hnRNP A1 also similarly obstructed the topo I revitalization. In addition, CPT has improved the topo I active inhibition to HshnRNP A1 under both existence.HshnRNP A1 or DmHrb87F united from the above-mentioned result with CPT, and the possibility of adjusting the topo I revitalization was suggested.