Pertussis toxin‐catalyzed ADP‐ribosylation of GTP‐binding proteins with digoxigenin‐conjugated NAD

FEBS Letters

Katada, Toshiaki¹  Takahashi, Katsunobu¹  Takei, Yoshinori¹  Kanaho, Yasunori¹ 

[1]Department of Life Science, Tokyo Institute of Technology, Yokohama 227, Japan

关键词: ADP-ribosylation;    Digoxigenin;    GTP-binding protein;    Pertussis toxin;    G protein;    GTP-binding protein;    Gs;    the stimulatory G protein of adenylyl cyclase;    Gi;    a family of homologous G proteins originally associated with inhibition of adenylyl cyclase;    Go;    a G protein purified from bovine brain;    DTT;    dithiothreitol;    IAP;    islet-activating protein (pertussis toxin);    SDS;    sodium dodecyl sulfate;    DIG-NAD;    digoxigenin-conjugated NAD;   

DOI  :  10.1016/0014-5793(94)80280-7

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

ADP-ribose moiety containing digoxigenin was transferred by pertussis toxin (IAP) to the α subunit of G_i (G_iα) from digoxigenin-conjugated NAD (DIG-NAD) in a βγ subunit-dependent manner. ADP-ribosylation of G_iα 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.

【 授权许可】

Unknown