Distinct kinetics of (H/K/N)Ras glucosylation and Rac1 glucosylation catalysed by Clostridium sordellii lethal toxin.
Mono-glucosylation of (H/K/N)Ras by Clostridium sordellii lethal toxin (TcsL) blocks critical survival signaling pathways, resulting in apoptotic cell death. One yet unsolved problem in studies on TcsL is the lack of a method allowing the specific detection of (H/K/N)Ras glucosylation. In this study, we identify the Ras(Mab 27H5) antibody as a glucosylation-sensitive antibody capable for the immunoblot detection of (H/K/N)Ras glucosylation in TcsL-treated cells. Alternative Ras antibodies including the K-Ras(Mab F234) antibody or the v-H-Ras(Mab Y13-159) antibody recognize Ras proteins regardless of glucosylation. (H/K)Ras are further shown to be more efficaciously glucosylated by TcsL than Rac1 in rat basophilic leukemia cells as well as in a cell-free system.
Huelsenbeck
Stefanie C
SC
Institut für Toxikologie, Medizinische Hochschule Hannover, D-30625 Hannover, Germany.
Klose
Ilona
I
Reichenbach
Maria
M
Huelsenbeck
Johannes
J
Genth
Harald
H
D000818:Animals; D000906:Antibodies; D017209:Apoptosis; D001427:Bacterial Toxins / Q000378:metabolism* / Q000494:pharmacology; D002384:Catalysis; D045744:Cell Line, Tumor; D006031:Glycosylation; D051381:Rats; D020830:rac1 GTP-Binding Protein / Q000276:immunology / Q000378:metabolism*; D018631:ras Proteins / Q000378:metabolism*
D000906:Antibodies; D001427:Bacterial Toxins; C072292:lethal toxin LT, Clostridium sordellii; D020830:rac1 GTP-Binding Protein; D018631:ras Proteins