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Pertussis toxin‐catalyzed ADP‐ribosylation of GTP‐binding proteins with digoxigenin‐conjugated NAD
Author(s) -
Takei Yoshinori,
Takahashi Katsunobu,
Kanaho Yasunori,
Katada Toshiaki
Publication year - 1994
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(94)80280-7
Subject(s) - nad+ kinase , adp ribosylation , digoxigenin , diphtheria toxin , pertussis toxin , biochemistry , microbiology and biotechnology , chemistry , biology , toxin , g protein , enzyme , receptor , gene expression , gene , in situ hybridization
ADP‐ribose moiety containing digoxigenin was transferred by pertussis toxin (IAP) to the α subunit of G i (G i α) from digoxigenin‐conjugated NAD (DIG‐NAD) in a βγ subunit‐dependent manner. ADP‐ribosylation of G i α with DIG‐NAD plus IAP was inhibited by native NAD. These results indicate that nonradiolabeled DIG‐NAD also serves as the substrate for IAP‐catalyzed ADP‐ribosylation of G proteins. Using DIG‐NAD and fluorescein isothiocyanate‐labeled anti‐digoxigenin antibody, IAP‐sensitive G protein(s) was found to be exist in nuclei as well as plasma membranes of rat liver and HeLa cells. Thus, DIG‐NAD is useful to identify pertussis toxin‐substrate G proteins.