Identification of rho as a substrate for botulinum toxin C3-catalyzed ADP-ribosylation.
Recombinant Aplysia rho and a GTP-binding protein purified from human neutrophil membranes (G22K) were ADP-ribosylated by botulinum toxin C3 with stoichiometries of 0.8 and 0.6, respectively. Rho and G22K appeared to be different proteins since (i) rho migrated faster on polyacrylamide gels, (ii) unlike G22K, rho did not require the presence of cytosol to be ADP-ribosylated, (iii) G22K was not recognized by an anti-rho antiserum, and (iv) antibody 142-24E05 recognized G22K effectively but only poorly cross reacted with rho. ADP-ribosylation had no effect on the ability of rho to bind or hydrolyse GTP. Therefore, it appears that there are multiple botulinum toxin C3 substrates and that the toxin exerts its effects on cell function by a mechanism other than modulating the GTPase activity of rho.
Quilliam
L A
LA
Department of Immunology, Research Institute of Scripps Clinic, La Jolla, CA 92037.
Lacal
J C
JC
Bokoch
G M
GM
D000246:Adenosine Diphosphate Ribose / Q000378:metabolism; D000818:Animals; D000911:Antibodies, Monoclonal; D001048:Aplysia; D001905:Botulinum Toxins / Q000378:metabolism*; D002417:Cattle; D002462:Cell Membrane / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel; D019204:GTP-Binding Proteins / Q000378:metabolism*; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D006160:Guanosine Triphosphate / Q000031:analogs & derivatives / Q000378:metabolism; D006801:Humans; D006868:Hydrolysis; D007700:Kinetics; D008565:Membrane Proteins / Q000378:metabolism*; D008970:Molecular Weight; D009504:Neutrophils / Q000032:analysis; D011994:Recombinant Proteins / Q000378:metabolism; D013873:Thionucleotides / Q000378:metabolism; D020743:rhoB GTP-Binding Protein
D000911:Antibodies, Monoclonal; D008565:Membrane Proteins; D011994:Recombinant Proteins; D013873:Thionucleotides; D000246:Adenosine Diphosphate Ribose; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D006160:Guanosine Triphosphate; D001905:Botulinum Toxins; D019204:GTP-Binding Proteins; D020743:rhoB GTP-Binding Protein