Pertussis toxin-mediated ADP-ribosylation of target proteins in Chinese hamster ovary cells involves a vesicle trafficking mechanism. Infect Immun 1995 Mar;63(3):825-32
Date
03/01/1995Pubmed ID
7868253Pubmed Central ID
PMC173077DOI
10.1128/iai.63.3.825-832.1995Scopus ID
2-s2.0-0028905448 (requires institutional sign-in at Scopus site) 46 CitationsAbstract
Pertussis toxin (PT)-catalyzed ADP-ribosylation of target proteins in intact Chinese hamster ovary (CHO) cells was evaluated with an in vitro ADP-ribosylation assay. In this assay, a postnuclear supernatant was prepared from CHO cells and used as a source of PT-sensitive target proteins for in vitro [32P[ADP-ribosylation. The postnuclear supernatant contained three proteins that were ADP-ribosylated in vitro, with apparent molecular masses of 50, 45, and 42 kDa. The 42- and 45-kDa proteins were membrane associated, while the 50-kDa protein was soluble. Following PT treatment of CHO cells, the 42- and 45-kDa proteins were not available for in vitro ADP-ribosylation, while the soluble 50-kDa protein remained available for in vitro ADP-ribosylation. The decrease in the availability of the 42- and 45-kDa proteins to in vitro ADP-ribosylation was proportional to the PT concentration and time of incubation with CHO cells. Western immunoblot analysis showed that extracts from PT-treated CHO cells and control CHO cells possessed equivalent amounts of two proteins that were recognized by anti-Gi protein antiserum. The two proteins recognized by anti-Gi protein antiserum from PT-treated cells migrated with higher apparent molecular weights than the two proteins from control cells. This was consistent with the in vivo ADP-ribosylation of the two proteins by PT.(ABSTRACT TRUNCATED AT 250 WORDS)
Author List
Xu Y, Barbieri JTAuthor
Joseph T. Barbieri PhD Professor in the Microbiology and Immunology department at Medical College of WisconsinMESH terms used to index this publication - Major topics in bold
Adenosine Diphosphate RiboseAmmonium Chloride
Animals
Biological Transport
Brefeldin A
CHO Cells
Cell Compartmentation
Cricetinae
Cyclopentanes
Detergents
GTP-Binding Proteins
Intracellular Membranes
Models, Biological
Pertussis Toxin
Protein Processing, Post-Translational
Protein Synthesis Inhibitors
Subcellular Fractions
Virulence Factors, Bordetella