Role of the A subunit of pertussis toxin in alteration of Chinese hamster ovary cell morphology.
The mechanism by which pertussis toxin induces morphological changes in Chinese hamster ovary cells was studied to determine whether the resulting clustered growth pattern is due to toxin-catalyzed ADP-ribosylation of a cellular substrate. While pertussis toxin was extremely potent in inducing morphological changes in Chinese hamster ovary cells, preparations of isolated A subunit or B oligomer exhibited greatly reduced activity. The clustered growth response of these cells correlated with ADP-ribosylation of a 41-kilodalton cellular substrate for the toxin in that the toxin concentration and time of exposure to the toxin required for ADP-ribosylation were the same as those needed for alterations in cellular morphology. Moreover, pertussis toxin modified by either chemical or photolytic methods exhibited similar decreases in the ability to ADP-ribosylate the cellular substrate and alter cell morphology. These results suggest that clustering of Chinese hamster ovary cells is due to toxin-catalyzed ADP-ribosylation of a 41-kilodalton substrate. Therefore, alteration in Chinese hamster ovary cell morphology can be used as a measure of toxin activity. This assay should prove to be a useful tool in the development and evaluation of new pertussis vaccines.
D000246:Adenosine Diphosphate Ribose / Q000502:physiology*; D000818:Animals; D002449:Cell Aggregation / Q000187:drug effects; D002478:Cells, Cultured / Q000166:cytology / Q000187:drug effects*; D006224:Cricetinae; D003412:Cricetulus; D005260:Female; D008970:Molecular Weight; D010053:Ovary; D037342:Pertussis Toxin; D011506:Proteins / Q000378:metabolism; D013997:Time Factors; D010566:Virulence Factors, Bordetella / Q000494:pharmacology*
D011506:Proteins; D010566:Virulence Factors, Bordetella; D000246:Adenosine Diphosphate Ribose; D037342:Pertussis Toxin