The accessibilities of histones in nucleosome cores to an arginine-specific protease.
Submaxillaris protease, reportedly specific for arginine residues, was used to probe accessible arginines of chicken erythrocyte nucleosome cores. The relative rates of digestion of histones in nucleosome cores by this protease were H3 greater than H2b greater than H4 much greater than H2a. At most, 8 of 52 total arginines among the four core histones were reasonably accessible to attack. Sites most rapidly cleaved were Arg-26 in H3, a site approximately 13 residues from the NH2 terminus of H2b, and Lys-12 or Arg-17 in H4. Five sites attacked more slowly were Arg-8, -128, or -129 and Arg-49, -52, or -53 in H3; Arg-3 and Arg-17 or -19 in H4; and a site near one terminus of H2b or H3. H2a and fragments resulting from the above cleavages were highly resistant to attack, even after prolonged incubation. Similar results were obtained upon digestion of histones in intact chromatin. H1 and H5 in whole chromatin were attacked at rates comparable to H3. Seven of the eight accessible sites lie outside of structure-forming histone sequences, i.e. sequences that are immobilized in histone complexes, indicating that these sequences are inaccessible in nucleosome cores. The single exceptional site noted, approximately 50 residues from the NH2 terminus of H3, is consistent with previous observations that Glu-51 and Glu-60 of H3 in nucleosomes are accessible to attack by S. aureus protease (Rill, R. L., and Oosterhof, D. K. (1981). J. Biol. Chem. 256, 12687-12691). The relationships of these protease accessibilities to NMR assignments of mobile histone tails in nucleosome cores are discussed.
D000818:Animals; D002467:Cell Nucleus / Q000032:analysis; D002645:Chickens; D010450:Endopeptidases / Q000378:metabolism*; D004912:Erythrocytes / Q000032:analysis; D006657:Histones / Q000097:blood* / Q000302:isolation & purification; D007700:Kinetics; D046911:Macromolecular Substances; D008970:Molecular Weight; D009707:Nucleosomes / Q000032:analysis*; D010446:Peptide Fragments / Q000032:analysis
D006657:Histones; D046911:Macromolecular Substances; D009707:Nucleosomes; D010446:Peptide Fragments; D010450:Endopeptidases; C034735:arginine protease