Multiple species and isoforms of Bordetella pertussis toxin substrates.

In purified G proteins from bovine brain cortex the ADP-ribosylated substrates of Bordetella pertussis toxin (PT) can be resolved in three polypeptides by polyacrylamide gel electrophoresis: a 39 kDa major substrate, corresponding to Go alpha and two others (40 and 41 kDa) assigned to alpha subunits of Gi-like proteins. These three polypeptides were also detected in membranes of normal cells or tissues from neuronal and endocrine origins. In contrast, in membranes from other origins, only two PT substrates at 41 and 40 kDa were resolved; the latter being the most abundant ADP-ribosylated substrate in human platelets and C6 glioma cells. In these cells, electrophoretic patterns of PT-radiolabeled proteolytic fragments derived from the 40 kDa peptide were different to those from the 39 and 41 kDa polypeptides of purified G proteins. However, isoelectrofocusing and two dimensional analyses showed that the 40 kDa and 39 kDa (but not the 41 kDa) PT substrate of purified G proteins exhibited similar isoforms.