已经在许多原核和真核物种和病毒中观察到ADP-核糖基转移酶活性,并且参与许多细胞过程。包括细胞信号,DNA修复,基因调控与细胞凋亡。在许多细菌毒素中,单ADP-核糖基转移酶是导致宿主细胞毒性的主要原因。已经使用了几种方法来分析这种生物系统,从测量其酶产物到其功能。通过使用单ADP-核糖结合蛋白,我们现在开发了一种ELISA方法来评估天然百日咳毒素单ADP-核糖基转移酶活性及其在百日咳疫苗中的残留活性。这种新方法在大多数实验室中易于执行且适应性强。理论上,该测定系统也非常通用,可以测量其他细菌如霍乱中的酶活性,梭菌属,大肠杆菌,白喉,百日咳,假单胞菌,沙门氏菌和葡萄球菌只需转换为各自的肽底物。此外,这种单ADP-核糖结合蛋白也可用于染色在凝胶或膜上分离的单ADP-核糖基产物。
ADP-ribosyltransferase activities have been observed in many prokaryotic and eukaryotic species and viruses and are involved in many cellular processes, including cell signalling, DNA repair, gene regulation and apoptosis. In a number of bacterial toxins, mono ADP-ribosyltransferase is the main cause of host cell cytotoxicity. Several approaches have been used to analyse this biological system from measuring its enzyme products to its functions. By using a mono ADP-ribose binding protein we have now developed an ELISA method to estimate native pertussis toxin mono ADP-ribosyltransferase activity and its residual activities in pertussis vaccines as an example. This new approach is easy to perform and adaptable in most laboratories. In theory, this assay system is also very versatile and could measure the enzyme activity in other bacteria such as Cholera, Clostridium, E. coli, Diphtheria, Pertussis, Pseudomonas, Salmonella and Staphylococcus by just switching to their respective peptide substrates. Furthermore, this mono ADP-ribose binding protein could also be used for staining mono ADP-ribosyl products resolved on gels or membranes.