title: Isolation and properties of the trypsin-derived ADP-ribosyl peptide from diphtheria toxin-modified yeast elongation factor 2.
abstract: We have developed a method for the purification in micromole amounts of the trypsin-derived ADP-ribosyl peptide from diphtheria toxin-modified yeast elongation factor 2 (EF-2). EF-2 was partially purified (15 to 20% purity) by ammonium sulfate precipitation and DEAE-Sephadex chromatography. After [3H]ADP-ribosylation by [3H]nad+ and diphtheria toxin, EF-2 was digested with trypsin and a homogeneous [3H]ADP-ribosyl peptide was isolated by chromatography on DEAE-Sephadex and dihydroxyboryl-substituted cellulose. Based on the amount of ADP-ribose acceptor activity in the crude extract, the overall yield of the peptide was 35%. The yeast peptide contains an unusual amino acid (X) which is the site of ADP ribosylation and is apparently identical to the amino acid reported from rat liver EF-2 by Robinson et al. (Robinson, E. A., Hendriksen, O., and Maxwell, E.S. (1974) J. Biol. Chem. 249, 5088-5093). The sequence of the 15-residue yeast peptide was determined to be: Val-Asn-Ile-Leu-Asp-Val-Thr-Leu-His-Ala-Asp-Ala-Ile-X-Arg. The 11 COOH-terminal residues of this peptide and of the homologous 15-residue peptide reported by Maxwell and co-workers from rat liver EF-2 are identical.
mesh_terms: D000247:Adenosine Diphosphate Sugars; D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D004167:Diphtheria Toxin; D009702:Nucleoside Diphosphate Sugars; D010445:Peptide Elongation Factors; D010455:Peptides / Q000302:isolation & purification; D012266:Ribose; D014357:Trypsin
chemical_list: D000247:Adenosine Diphosphate Sugars; D000596:Amino Acids; D004167:Diphtheria Toxin; D009702:Nucleoside Diphosphate Sugars; D010445:Peptide Elongation Factors; D010455:Peptides; D012266:Ribose; D014357:Trypsin