title: Site-specificity of histone H1 methylation by two H1-specific protein-lysine N-methyltransferases from Euglena gracilis.
abstract: 1. The histone H1 fractions from rat spleen and liver were used as substrates for two H1-specific protein-lysine N-methyltransferases, V-A and V-B (protein methylase III) from Euglena gracilis. 2. When the enzymatically [methyl-3H]labeled H1 fractions were resolved by two-dimensional gel electrophoresis, four subtypes were found to be methylated (H1b, H1c, H1d and H1e). Both enzymes methylated H1c and H1b to approximately the same extent; H1d and H1e were methylated preferentially by enzyme V-B and V-A, respectively. 3. Histone H1c, [methyl-3H]labeled by the methyltransferase V-A, which had been digested by arginine-specific protease (Arg C protease), showed a single radioactive peptide on HPLC, indicating methylation site specificity of the enzyme. 4. Arg C protease-digestion of [methyl-3H]labeled H1c labeled by methyltransferase V-B indicated that this enzyme methylated two sites on the histone molecule. 5. The histone H1c methylation sites of these two enzymes did not overlap, indicating the two enzymes have different site specificity. 6. In combination with the other results, this suggests that the two enzymes serve discrete purposes, possibly involving the presumed different actions of histone H1 subtypes.
authors.affiliation: Fels Institute for Cancer Research and Molecular Biology, Temple University School of Medicine, Philadelphia, PA 19140.
authors.forename: B F
authors.initials: BF
authors.lastname: Frost
authors.forename: K S
authors.initials: KS
authors.lastname: Park
authors.forename: M
authors.initials: M
authors.lastname: Tuck
authors.forename: S
authors.initials: S
authors.lastname: Disa
authors.forename: S
authors.initials: S
authors.lastname: Kim
authors.forename: W K
authors.initials: WK
authors.lastname: Paik
mesh_terms: D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001665:Binding Sites; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D015180:Electrophoresis, Gel, Two-Dimensional; D010450:Endopeptidases; D005056:Euglena gracilis / Q000201:enzymology*; D011495:Histone-Lysine N-Methyltransferase / Q000378:metabolism*; D006657:Histones / Q000378:metabolism*; D008099:Liver / Q000378:metabolism; D008745:Methylation; D010446:Peptide Fragments / Q000302:isolation & purification; D011496:Protein Methyltransferases / Q000378:metabolism*; D051381:Rats; D012697:Serine Endopeptidases; D013154:Spleen / Q000378:metabolism
chemical_list: D000596:Amino Acids; D006657:Histones; D010446:Peptide Fragments; D011496:Protein Methyltransferases; D011495:Histone-Lysine N-Methyltransferase; D010450:Endopeptidases; C044499:submandibular proteinase A; D012697:Serine Endopeptidases