Production of an antiserum specific to the ADP‐ribosylated form of elongation factor 2 from archaebacteria and eukaryotes

FEBS Letters

Siegmund, Klaus-Detlef¹  Klink, Friedrich¹ 

[1] Biochemisches Institut der Christian-Albrechts Universität, Olshausenstr. 40-60, D-2300 Kiel 1, Germany

关键词: Antiserum;    Elongation factor 2;    ADP-ribosylation;    Sulfolobus acidocaldarius;    EF-2;    elongation factor 2;    ADPR-EF-2;    ADP-ribosylated elongation factor 2;    PBS;    phosphate buffered saline;    rEF-2;    recombinant elongation factor 2;   

DOI  :  10.1016/0014-5793(92)80921-3

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

An antiserum to ADP-ribosylated elongation factor 2 (ADPR-EF-2) from S. acidocaldarius was raised in rabbits using stained, homogenized, ADPR-EF-2-containing slices from SDS-gels as a source of antigen. Elongation factor 2 (EF-2) from S. acidocaldarius was cloned in E. coli and the expressed gene product was used in order to adsorb all anti-EF-2 antibodies which do not contain the ADP-ribosyl group within their epitopes, as E. coli is unable to synthesize the ADP-ribosyl acceptor diphthamide. The remaining antibodies were specific to ADP-ribosylated EF-2 from Thermoplasma acidophilum, S. acidocaldarius and Desulfurococcus mucosus. ADP-ribosylated EF-2 from eukaryotic sources also reacted with the adsorbed antiserum as shown for EF-2 isolated from the killi-fish Cynolebias whitei, the mouse species BALB/c and Han/Wistar rats. The adsorbed antiserum did not cross-react with ADP-ribosylated actin or rho protein or with FAD-containing D-amino acid oxidase.

【 授权许可】

Unknown