title: Identification of the phosphorylation sites of H2B histone by a catalytic fragment of p72syk from porcine spleen.
abstract: Phosphorylated sites of calf thymus H2B histone were investigated with a catalytic fragment of 72 kDa protein-tyrosine kinase (p72syk). Three of five tyrosine residues in H2B histone can be phosphorylated by this kinase. In this analysis, H2B histone was thoroughly phosphorylated in vitro with [gamma-32P]ATP and the kinase, and then digested with a lysylendopeptidase. The resulting radioactive phosphopeptides were separated by a reverse-phase column on high performance liquid chromatography. Subsequent sequential Edman degradation of the purified phosphopeptides revealed that 40Y, 83Y and 121Y were phosphorylated. 121Y is the major phosphorylated residue in H2B histone. No phosphorylation was detected in 37Y and 42Y. Although the consensus sequence was not defined from these analyses, our data suggest that higher-order structure(s) in addition to primary one may participate in recognition of H2B histone by this protein kinase.
authors.affiliation: Department of Biochemistry, Fukui Medical School, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Sakai
authors.forename: Y
authors.initials: Y
authors.lastname: Tanaka
authors.forename: M
authors.initials: M
authors.lastname: Asahi
authors.forename: R
authors.initials: R
authors.lastname: Shimomura
authors.forename: T
authors.initials: T
authors.lastname: Taniguchi
authors.forename: E
authors.initials: E
authors.lastname: Hashimoto
authors.forename: H
authors.initials: H
authors.lastname: Yamamura
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D004792:Enzyme Precursors / Q000378:metabolism*; D006657:Histones / Q000235:genetics / Q000378:metabolism*; D047908:Intracellular Signaling Peptides and Proteins; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry / Q000302:isolation & purification; D010748:Phosphopeptides / Q000737:chemistry / Q000302:isolation & purification; D010766:Phosphorylation; D011505:Protein-Tyrosine Kinases / Q000378:metabolism*; D013154:Spleen / Q000201:enzymology*; D013552:Swine; D000072377:Syk Kinase
chemical_list: D004792:Enzyme Precursors; D006657:Histones; D047908:Intracellular Signaling Peptides and Proteins; D010446:Peptide Fragments; D010748:Phosphopeptides; D011505:Protein-Tyrosine Kinases; D000072377:Syk Kinase