Estimating the amounts of ADP-ribosylatable active elongation factor-2 in mammalian cell-free extracts.
The content of the elongation factor (EF-2) can be measured by diphtheria toxin-dependent ADP-ribosylation in cell-free extracts of samples prepared from small amounts of tissues and cells containing less than 100 micrograms of total protein. A 20 min in vitro assay, in which a radioactive ADP-ribosyl residue is transferred specifically and 1:1 stoichiometrically to EF-2, is sufficient to estimate the total amounts of ADP-ribosylatable active EF-2. The method is very useful for monitoring changing levels of EF-2 during various pathological and biological processes, including cell cycle, ageing, cancer and other diseases.
Riis
B
B
Department of Chemistry, Aarhus University, Denmark.
Rattan
S I
SI
Clark
B F
BF
D000246:Adenosine Diphosphate Ribose / Q000378:metabolism*; D000818:Animals; D002250:Carbon Radioisotopes; D004167:Diphtheria Toxin / Q000494:pharmacology; D006367:HeLa Cells / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D008099:Liver / Q000378:metabolism*; D009243:NAD / Q000378:metabolism*; D020652:Peptide Elongation Factor 2; D010445:Peptide Elongation Factors / Q000032:analysis / Q000378:metabolism*; D010750:Phosphoproteins / Q000378:metabolism*; D011865:Radioisotope Dilution Technique; D051381:Rats
D002250:Carbon Radioisotopes; D004167:Diphtheria Toxin; D020652:Peptide Elongation Factor 2; D010445:Peptide Elongation Factors; D010750:Phosphoproteins; D009243:NAD; D000246:Adenosine Diphosphate Ribose