Conformation and domain structure of the non-histone chromosomal proteins, HMG 1 and 2. Isolation of two folded fragments from HMG 1 and 2.

Proteins HMG 1 and 2 have been digested with trypsin and two major products, stable to further digestion between 8 min and 2 h, have been purified (peptides A and B). Peptide B from HMG 1 has been identified as residues 12-75 and peptide A as residues 94/96-169 by amino acid analyses and Edman degradations. Peptide B spontaneously folds with the formation of 51% helix and exhibits the majority of the perturbed NMR resonances characteristic of folded intact HMG 1. Peptide B is stably folded in the presence of 150 mM NaCl between pH 3 and 10, like intact HMG 1. Peptide A forms 30% alpha-helix and also exhibits tertiary folding but is denatured by pH 10. The 11 N-terminal residues removed by trypsin contain both sites of post-synthetic acetylation (residues 2 and 11), a situation very similar to that found with core histones. It is proposed that HMG 1 and 2 consist of four structural domains, viz: (a) residues 1-11, (b) residues 12 to approximately 75, (c) residues 94-169 and (d) the very acidic region beyond residue 169. The instability of peptide A may mean that it is not a truly independent domain. No structural similarities to histone H1 are therefore observed in HMG 1 and 2.