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Production of an antiserum specific to the ADP‐ribosylated form of elongation factor 2 from archaebacteria and eukaryotes
Author(s) -
Siegmund Klaus-Detlef,
Klink Friedrich
Publication year - 1992
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(92)80921-3
Subject(s) - antiserum , elongation factor , biology , adp ribosylation , biochemistry , sulfolobus acidocaldarius , microbiology and biotechnology , antigen , gene , enzyme , ribosome , nad+ kinase , genetics , rna , archaea
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.