title: Purification of histone H1 polypeptides by high-performance cation-exchange chromatography.
abstract: Calf thymus histone 1 (H1) was cleaved by chemical and enzymatic methods and the resulting polypeptides were fractionated by high-performance cation-exchange. Up to 1 mg of H1 polypeptides were loaded onto a 50 x 5 mm I.D. cation-exchange column and fractionated to greater than 95% purity in less than 30 min. This is the first report on the separation of H1 polypeptides by a strong cation-exchange matrix. In addition, the high-performance cation-exchange chromatography protocol represents a significant decrease in fractionation time when compared to conventional ion-exchange and gel filtration chromatography. The utility of this procedure is shown when the H1 peptides purified by the protocol were used to define antigenic domains of H1 band by procainamide-induced lupus and idiopathic systemic lupus erythematosus. The majority of the sera tested by enzyme-linked immunoassay (ELISA) reacted to the C-terminal peptides of H1 indicating this to be the major antigenic domain of H1.
authors.affiliation: Joint Injury and Diseases Research Group, Faculty of Medicine, University of Calgary, Alberta, Canada.
authors.forename: J
authors.initials: J
authors.lastname: Gohill
authors.forename: J D
authors.initials: JD
authors.lastname: Pauls
authors.forename: M J
authors.initials: MJ
authors.lastname: Fritzler
mesh_terms: D000818:Animals; D001974:Bromosuccinimide / Q000494:pharmacology; D002417:Cattle; D002853:Chromatography, Liquid / Q000379:methods*; D002918:Chymotrypsin / Q000494:pharmacology; D004797:Enzyme-Linked Immunosorbent Assay; D006657:Histones / Q000032:analysis / Q000187:drug effects / Q000276:immunology / Q000302:isolation & purification*; D008180:Lupus Erythematosus, Systemic / Q000097:blood / Q000276:immunology; D011342:Procainamide; D012965:Sodium Chloride; D013917:Thrombin / Q000494:pharmacology; D013950:Thymus Gland / Q000032:analysis
chemical_list: D006657:Histones; D012965:Sodium Chloride; D002918:Chymotrypsin; D013917:Thrombin; D001974:Bromosuccinimide; D011342:Procainamide