title: ADP-ribosylation of elongation factor 2 by diphtheria toxin. Isolation and properties of the novel ribosyl-amino acid and its hydrolysis products.
abstract: We have isolated the novel amino acid, amino acid X, in elongation factor 2 (EF-2) which is ADP-ribosylated by diphtheria toxin (Robinson, E. A., Hendriksen, O., and Maxwell, E. S. (1974) J. Biol. Chem. 249, 5088-5093). The pure ribosyl-amino acid was obtained in milligram quantities following sequential enzymatic degradation of yeast ADP-ribosyl-EF-2 (Van Ness, B. G., Howard, J. B., and Bodley, J. W. (1978) J. Biol. Chem. 253, 8687-8690) with an overall yield of 30 to 60%. In the course of acid hydrolysis, the ribosyl-amino acid was converted first to an intermediate compound which did not contain ribose. Further acid hydrolysis converted the intermediate to amino acid X and ammonia. Alkaline hydrolysis of the ribosyl-amino acid produced a different intermediate which contained ribose. We have isolated the two hydrolysis intermediates and on the basis of the properties of all four compounds we conclude that the ribosyl-amino acid contains an amido function which is not involved in a glycosidic linkage and a net positive charge of 2 at acid pH. We believe that the amino acid amide occurs in EF-2 and for this compound we propose the trivial name diphthamide. Acid hydrolysis of diphthamide yields amino acid X for which we propose the trivial name diphthine.
mesh_terms: D000246:Adenosine Diphosphate Ribose / Q000378:metabolism*; D004167:Diphtheria Toxin / Q000378:metabolism*; D006639:Histidine / Q000031:analogs & derivatives* / Q000032:analysis; D006868:Hydrolysis; D007700:Kinetics; D009702:Nucleoside Diphosphate Sugars / Q000378:metabolism*; D020652:Peptide Elongation Factor 2; D010445:Peptide Elongation Factors / Q000378:metabolism*; D012263:Ribonucleosides / Q000032:analysis*; D012441:Saccharomyces cerevisiae / Q000378:metabolism
chemical_list: D004167:Diphtheria Toxin; D009702:Nucleoside Diphosphate Sugars; D020652:Peptide Elongation Factor 2; D010445:Peptide Elongation Factors; D012263:Ribonucleosides; D000246:Adenosine Diphosphate Ribose; D006639:Histidine; C027529:diphthine; C027528:ribosyl-diphthamide