title: Isolation and characterization of folded fragments released by Staphylococcal aureus proteinase from the non-histone chromosomal protein HMG-1.
abstract: HMG-1 was isolated from newborn calf thymus without exposure to overt denaturing conditions. The purified protein was digested under several solvent conditions with the proteinase (endoproteinase GluC) from Staphylococcus aureus strain V8. We found that the preferred site of attack by the enzyme on HMG-1 was influenced markedly by ionic strength and temperature. In 0.35 M NaCl/50 mM Tris-phosphate (pH 7.8) at 37 degrees C, cleavage near the junction between the A and B domains is predominant, as previously reported by Carballo et al. (EMBO J. 2 (1983) 1759-1764). However, in 50 mM Tris-phosphate (pH 7.8) lacking NaCl and at 0 degrees C, cleavage between the B and C domains strongly predominates. Three major products of the digestions were purified and characterized. The fragment consisting of domains B and C was found by circular dichroism to contain a substantial amount of helix. This re-emphasizes the importance of avoiding overt denaturing conditions when working with members of the HMG-1 family.
authors.affiliation: Department of Biochemistry, Kansas State University, Manhattan 66506.
authors.forename: K K
authors.initials: KK
authors.lastname: Abdul-Razzak
authors.forename: M L
authors.initials: ML
authors.lastname: Denton
authors.forename: D J
authors.initials: DJ
authors.lastname: Cox
authors.forename: G R
authors.initials: GR
authors.lastname: Reeck
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D002942:Circular Dichroism; D006609:High Mobility Group Proteins / Q000032:analysis* / Q000648:ultrastructure; D008666:Metalloendopeptidases / Q000494:pharmacology; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010446:Peptide Fragments / Q000032:analysis / Q000302:isolation & purification; D011487:Protein Conformation; D012492:Salts / Q000494:pharmacology; D013379:Substrate Specificity; D013696:Temperature
chemical_list: D006609:High Mobility Group Proteins; D010446:Peptide Fragments; D012492:Salts; D008666:Metalloendopeptidases; C022653:auR protein, Staphylococcus aureus