In vivo pertussis toxin administration: effects on the function and levels of Gi alpha proteins and their messenger ribonucleic acids

Endocrinology
V Ramkumar, G L Stiles

Abstract

Pertussis toxin ADP-ribosylates and functionally inactivates a number of guanine nucleotide-binding proteins (G-proteins), including the inhibitory G-protein, Gi. Once ADP-ribosylated, the subsequent duration of inactivation and fate of these proteins in cells are unknown. In the present study, therefore, we have attempted to answer some of these questions in an in vivo rat adipocyte model. Our results indicate a rather prolonged (greater than 2 weeks) inactivation of Gi, as evidenced by loss of R-phenylisopropyladenosine-mediated inhibition of adenylate cyclase activity in rat adipocyte membranes after administration of the toxin in vivo. This was associated with a complete loss of pertussis toxin ADP-ribosylatable substrates on days 3 and 15, with only partial recovery by day 28 after initiation of treatment. Specific antibodies used to quantitate the levels of alpha i1- and alpha 12-subunits of alpha i indicate a 2-fold increase in the levels of these proteins by day 3, followed by a progressive decrease and subsequent recovery in these subunits to 53%, 47%, and 87% of the control value on days 15, 20, and 28, respectively. No change in the levels of alpha s was observed at all time points tested. The levels of mRNA encoding...Continue Reading

Citations

Oct 1, 1994·The Journal of Clinical Investigation·M G DaviesP O Hagen
Mar 1, 1992·Journal of Neurochemistry·J D SteketeeP W Kalivas
Apr 5, 1991·Brain Research·J D SteketeeP W Kalivas

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