Chemical and enzymatic analysis of covalent bonds between peptides and chromosomal DNA.
DNA from Ehrlich ascites tumor (EAT) cells and from human placenta was examined for covalent bonds between hydroxy amino acid residues in peptides and nucleotide phosphate groups. The residual proteinaceous material in highly purified DNA was radiolabelled with 125Iodine and the linking-groups between peptides and nucleotides released by combined protease and nuclease treatment were investigated with respect to their chemical and enzymatic stabilities. The residual nucleotide(s)-peptide(s) fraction from DNA isolated after prolonged alkaline cell lysis and phenol extraction contains mainly alkali and acid-stable but phosphodiesterase-sensitive peptide-nucleotide complexes which indicates phosphodiesters between tyrosyl residues in peptides and nucleotide phosphates. In contrast, the linking-group fraction from DNA isolated under native conditions contains additional peptide components. (a) Phospho-peptides that co-purify with DNA but that are not covalently bound to nucleotides. (b) A fraction of peptides that is released from nucleotides by alkali in a time and concentration-dependent reaction. Evidence is presented indicating that the latter fraction involves phospho-triesters between hydroxy amino acid residues in peptides and internucleotide phosphates. The phosphodiesters between hydroxy amino acids and nucleotide phosphates representing the predominant class of peptide-nucleotide complexes in alkali-denatured DNA are most likely side products of peptide-nucleotide phospho-triester hydrolysis.
Juodka
B
B
Department of Biochemistry and Biophysics, Vilnius University, Lithuania.
Pfütz
M
M
Werner
D
D
D000596:Amino Acids / Q000737:chemistry / Q000378:metabolism; D000818:Animals; D043303:Aryldialkylphosphatase; D002286:Carcinoma, Ehrlich Tumor; D002855:Chromatography, Thin Layer; D003198:Computer Simulation; D004247:DNA / Q000737:chemistry* / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008958:Models, Molecular; D010744:Phosphoric Monoester Hydrolases / Q000378:metabolism; D014407:Tumor Cells, Cultured
D000596:Amino Acids; D004268:DNA-Binding Proteins; D004247:DNA; D010744:Phosphoric Monoester Hydrolases; D043303:Aryldialkylphosphatase