PubMed1990
Certain microbial toxins ADP-ribosylate G proteins that may be related to those postulated to participate in secretion, whilst botulinum neurotoxins, produced by Clostridium botulinum, block Ca2(+)-dependent neurotransmitter release. Thus, botulinum, pertussis and cholera toxins were examined for ADP-ribosyl transferase activity using isolated nerve terminals. Although type D botulinum, cholera and pertussis toxins exhibited such enzymic activity, this was not detectable with types A or B botulinum neurotoxins or their individual chains, in any synaptosomal fraction. Botulinum type D and pertussis toxins ADP-ribosylated proteins with mol. wt approximately 24,000 and 42,000 respectively, whereas cholera toxin modified several proteins including a 51,000/47,000 mol. wt doublet. Pre-incubation of synaptosomes with type A, B or D toxins did not inhibit type D-induced labelling in the corresponding lysate. Similar pre-incubations with cholera or pertussis toxins reduced ADP-ribosylation of their substrates. Hence, under conditions in which these botulinum toxins were shown to block Ca2(+)-dependent transmitter release no ADP-ribosylated substrate was produced in the intact nerve terminals. Moreover, direct correlation was not found between the concentration dependencies of type D toxin for protein modification and inhibition of [3H]noradrenaline release from synaptosomes. These collective findings implicate C3, a non-neurotoxic contaminant of type D, in the enzymic action. The substrate for type D toxin was found in the cytosolic fraction and to a lesser extent in synaptic membranes, the reverse of the situation for pertussis toxin. A combination of the membranes and cytosol was required for maximal labelling of the 51,000/47,000 doublet by cholera toxin. Purified synaptic vesicles contained proteins labelled by type D and pertussis toxins but lacked major cholera toxin substrates. Future research will determine the possible involvement of these toxin-susceptible vesicular proteins in transmitter release.
Adenosine Diphosphate Ribose↗