PT  - JOURNAL ARTICLE
AU  - R Graf
AU  - J Codina
AU  - L Birnbaumer
TI  - Peptide inhibitors of ADP-ribosylation by pertussis toxin are substrates with affinities comparable to those of the trimeric GTP-binding proteins.
DP  - 1992 Nov 01
TA  - Molecular Pharmacology
PG  - 760--764
VI  - 42
IP  - 5
4099  - http://molpharm.aspetjournals.org/content/42/5/760.short
4100  - http://molpharm.aspetjournals.org/content/42/5/760.full
SO  - Mol Pharmacol1992 Nov 01; 42
AB  - Pertussis toxin (PTX) ADP-ribosylates alpha subunits of GTP-binding proteins (G proteins) when they are in association with beta gamma dimers, and free alpha subunits are thought not to be substrates under standard assay conditions. We now report the rather unexpected discovery that synthetic peptides encompassing the last 10-20 amino acids of alpha subunits of PTX-sensitive G proteins are substrates for PTX by themselves and in the absence of beta gamma dimers. As determined for G13, the Km of PTX for the 20-amino acid carboxyl-terminal peptide is 10-fold higher than that for the trimeric G protein. Interestingly, PTX ADP-ribosylates the free full length alpha 13 subunit with a Km not different from that of the trimer but with a Vmax that is only 1% of that with which it ADP-ribosylates the trimer. It follows that the primary role of beta gamma dimers in ADP-ribosylation of G proteins is one of increasing the Vmax of the reaction without affecting the Km of the substrate for the toxin. Mutant peptides lacking the ADP-ribose acceptor site act as competitive inhibitors.