Endogenous ADP-ribosylation of elongation factor 2 in polyoma virus-transformed baby hamster kidney cells.
- 1 January 1989
- journal article
- research article
- Published by Proceedings of the National Academy of Sciences in Proceedings of the National Academy of Sciences
- Vol. 86 (2), 554-557
- https://doi.org/10.1073/pnas.86.2.554
Abstract
Polyoma virus-transformed baby hamster kidney (pyBHK) cells were cultured in medium containing [32P]orthophosphate and 10% (vol/vol) fetal bovine serum. A 32P-labeled protein with an apparent molecular mass of 97 kDa was immunoprecipitated from cell lysates with antiserum to ADP-ribosylated elongation factor 2 (EF-2). The 32P labeling of the protein was enhanced by culturing cells in medium containing 2% serum instead of 10% serum. The 32P label was completely removed from the protein by treatment with snake venom phosphodiesterase and the digestion product was identified as [32P]AMP, indicating the protein was mono-ADP-ribosylated. HPLC analysis of tryptic peptides of the 32P-labeled 97-kDa protein and purified EF-2, which was ADP-ribosylated in vitro with diphtheria toxin fragment A and [32P]NAD, demonstrated an identical labeled peptide in the two proteins. The data strongly suggest that EF-2 was endogenously ADP-ribosylated in pyBHK cells. Maximum incorporation of radioactivity in EF-2 occurred by 12 hr and remained constant over the subsequent 12 hr. It was estimated that 30-35% of the EF-2 was ADP-ribosylated in cells cultured in medium containing 2% serum. When 32P-labeled cultures were incubated in medium containing unlabeled phosphate, the 32P label was lost from the EF-2 within 30 min.This publication has 31 references indexed in Scilit:
- On the nature of cellular ADP-ribosyltransferase from rat liver specific for elongation factor 2Biochemical and Biophysical Research Communications, 1986
- Endogenous ADP‐ribosylation of elongation factor 2 in polyribosome fraction of rabbit reticulocytesFEBS Letters, 1984
- ADP‐ribosyltransferase from beef liver which ADP‐ribosylates elongation factor‐2FEBS Letters, 1984
- Purification and Properties of an Altered Form of Elongation Factor 2 from Mutant Cells Resistant to Intoxication by Diphtheria ToxinEuropean Journal of Biochemistry, 1983
- Recognition of elongation factor 2 by diphtheria toxin is not solely defined by the presence of diphthamideFEBS Letters, 1980
- Primary structure at the site in beef and wheat elongation factor 2 of ADP‐ribosylation by diphtheria toxinFEBS Letters, 1979
- The regulation of protein synthesis in animal cells by serum factorsBiochemistry, 1976
- Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4Nature, 1970
- Diphtheria Toxin Submit Active in vitroScience, 1969
- Arrested Protein Synthesis in Polysomes of Cultured Chick Embryo CellsScience, 1966