title: The use of ubiquitin-peptide extensions as protein kinase substrates.
abstract: Four ubiquitin-peptide extensions prepared as cloned products in E. coli were tested as casein kinase II substrates. Two extensions containing the sequence Ser-Glu-Glu-Glu-Glu-Glu were readily phosphorylated by partially purified rabbit reticulocyte casein kinase II. The other two fusion proteins, which lack a consensus phosphorylation site for casein kinase II, did not serve as substrates under identical reaction conditions. Native ubiquitin was not phosphorylated by reticulocyte casein kinase II, nor have we observed its phosphorylation in crude extracts from HeLa cells, mouse liver, or Xenopus eggs. Ubiquitin's apparent lack of phosphorylatable residues coupled with its remarkable heat stability and rapid migration on sodium dodecyl sulfate-polyacrylamide gel electrophoresis gels make the protein an attractive carrier for carboxyl-terminal peptides containing specific phosphorylation sites. Such ubiquitin extension proteins should prove valuable as protein kinase substrates.
authors.affiliation: Department of Biochemistry, University of Utah, Salt Lake City 84132.
authors.forename: Y
authors.initials: Y
authors.lastname: Yoo
authors.forename: M
authors.initials: M
authors.lastname: Rechsteiner
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D047388:Casein Kinases; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D006367:HeLa Cells; D006801:Humans; D007700:Kinetics; D008099:Liver / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000378:metabolism*; D010766:Phosphorylation; D011494:Protein Kinases / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism*; D012156:Reticulocytes / Q000201:enzymology; D013379:Substrate Specificity; D014452:Ubiquitins / Q000378:metabolism*; D014981:Xenopus
chemical_list: D009842:Oligopeptides; D011994:Recombinant Proteins; D014452:Ubiquitins; D011494:Protein Kinases; D047388:Casein Kinases