Immunochemical detection of guanine nucleotide binding proteins mono-ADP-ribosylated by bacterial toxins.
Rabbits immunized with ADP-ribose chemically conjugated to carrier proteins developed antibodies reactive against guanine nucleotide binding proteins (G proteins) that had been mono-ADP-ribosylated by bacterial toxins. Antibody reactivity on immunoblots was strictly dependent on incubation of substrate proteins with both toxin and NAD and was quantitatively related to the extent of ADP-ribosylation. Gi, Go, and transducin (ADP-ribosylated by pertussis toxin) and elongation factor II (EF-II) (ADP-ribosylated by pseudomonas exotoxin) all reacted with ADP-ribose antibodies. ADP-ribose antibodies detected the ADP-ribosylation of an approximately 40-kilodalton (kDa) membrane protein related to Gi in intact human neutrophils incubated with pertussis toxin and the ADP-ribosylation of an approximately 90-kDa cytosolic protein, presumably EF-II, in intact HUT-102 cells incubated with pseudomonas exotoxin. ADP-ribose antibodies represent a novel tool for the identification and study of G proteins and other substrates for bacterial toxin ADP-ribosylation.
D036002:ADP Ribose Transferases; D000246:Adenosine Diphosphate Ribose / Q000032:analysis* / Q000276:immunology / Q000378:metabolism; D000906:Antibodies; D000936:Antigen-Antibody Complex; D001427:Bacterial Toxins; D002460:Cell Line; D017977:Deltaretrovirus; D005098:Exotoxins / Q000378:metabolism*; D019204:GTP-Binding Proteins / Q000032:analysis* / Q000276:immunology / Q000378:metabolism; D006801:Humans; D007938:Leukemia; D009504:Neutrophils / Q000502:physiology; D037342:Pertussis Toxin; D037521:Virulence Factors; D010566:Virulence Factors, Bordetella / Q000378:metabolism*; D000097668:Pseudomonas aeruginosa Exotoxin A
D036002:ADP Ribose Transferases; D000246:Adenosine Diphosphate Ribose; D000906:Antibodies; D000936:Antigen-Antibody Complex; D001427:Bacterial Toxins; D005098:Exotoxins; D019204:GTP-Binding Proteins; D037342:Pertussis Toxin; D037521:Virulence Factors; D010566:Virulence Factors, Bordetella; D000097668:Pseudomonas aeruginosa Exotoxin A