Estimating the amounts of ADP-ribosylatable active elongation factor-2 in mammalian cell-free extracts.

Abstract

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.

• Authors: B Riis, S I Rattan, B F Clark
• Journal: Journal of biochemical and biophysical methods (J Biochem Biophys Methods) Vol. 19 Issue 4 Pg. 319-25 (Oct 1989) ISSN: 0165-022X [Print] Netherlands
• PMID: 2693515 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Carbon Radioisotopes
• Diphtheria Toxin
• Peptide Elongation Factor 2
• Peptide Elongation Factors
• Phosphoproteins
• NAD
• Adenosine Diphosphate Ribose

Topics

• Adenosine Diphosphate Ribose (metabolism)
• Animals
• Carbon Radioisotopes
• Diphtheria Toxin (pharmacology)
• HeLa Cells (metabolism)
• Humans
• Kinetics
• Liver (metabolism)
• NAD (metabolism)
• Peptide Elongation Factor 2
• Peptide Elongation Factors (analysis, metabolism)
• Phosphoproteins (metabolism)
• Radioisotope Dilution Technique
• Rats