Production of an antiserum specific to the ADP-ribosylated form of elongation factor 2 from archaebacteria and eukaryotes.
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.
Siegmund
K D
KD
Biochemisches Institut, Christian-Albrechts Universität, Kiel, Germany.
Klink
F
F
D000244:Adenosine Diphosphate / Q000378:metabolism; D000818:Animals; D000907:Antibodies, Bacterial / Q000276:immunology*; D000918:Antibody Specificity; D003001:Cloning, Molecular; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay; D007106:Immune Sera / Q000276:immunology*; D007114:Immunization; D007525:Isoelectric Focusing; D020652:Peptide Elongation Factor 2; D010445:Peptide Elongation Factors / Q000276:immunology* / Q000378:metabolism; D011817:Rabbits; D051381:Rats; D012266:Ribose / Q000378:metabolism; D016945:Sulfolobus acidocaldarius / Q000276:immunology*
D000907:Antibodies, Bacterial; D007106:Immune Sera; D020652:Peptide Elongation Factor 2; D010445:Peptide Elongation Factors; D000244:Adenosine Diphosphate; D012266:Ribose