Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
7557119 | Analytical Biochemistry | 2018 | 5 Pages |
Abstract
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.
Keywords
NaADOVALPCHRPTBSDTTPTDABTSADPmAbMacro domainPTXPBSPRNIPVFHABSAFilamentous haemagglutininHydrogen peroxideadenosine diphosphatebovine serum albuminMonoclonal antibodyOvalbuminTris buffer salineEnzyme-linked immunosorbent assayELISAPertussis toxoiddiphtheria toxoidtetanus toxoidHISdithiothreitolpertussis toxinshdFormaldehydePhosphate buffered salineLysophosphatidylcholineNADHaemophilus influenzae type bnicotinamide adenine dinucleotidenicotinic acid adenine dinucleotideHibH2O2HisthistidinePertussis vaccinesInactivated poliovirus vaccineHorseradish peroxidasePertactinoptical densityhigh performance liquid chromatographyHPLC
Related Topics
Physical Sciences and Engineering
Chemistry
Analytical Chemistry
Authors
Catpagavalli Asokanathan, Sharon Tierney, Christina R. Ball, George Buckle, Ami Day, Simon Tanley, Adrian Bristow, Kevin Markey, Dorothy Xing, Chun-Ting Yuen,