title: Investigations of primary and secondary structure of porcine ubiquitin. Its N epsilon-acetylated lysine derivative.
abstract: N epsilon-acetylation in vitro of internal lysyl residues of Ub by p-nitro-phenyl acetate at pH 8.0 was performed. The position of acetylation sites are determined. (e.g. Fully acetylated: Lys-6, Lys-11 and Lys-33; partially free internal lysines: Lys-27, Lys-29; Lys-48 and probably Lys-63.) 55 cycles Edman degradation were performed and the first 53 N-terminal residues were identified. Secondary structural studies of ubiquitin have been carried out using the circular dichroism (CD) technique. No changes are noted upon heating to 100 degrees C at neutral pH even in the presence of 8 M urea but in 6 M guanidine-HCl extensive modification results. Ubiquitin with an average of 4.4 of its 7 lysines in the N epsilon-acetyl form shows little deviation from native protein. After reduction with dithiothreitol and subsequent removal of the mercaptan, significant changes in the secondary structure are noted. Circular dichroic measurements of ubiquitin indicated an alpha-helical content of about 10% whereas the secondary structural predictions of Chou and Fasman suggest a level of about 45%.
mesh_terms: D000107:Acetylation; D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D002942:Circular Dichroism; D004229:Dithiothreitol; D004912:Erythrocytes / Q000378:metabolism*; D019791:Guanidine; D006146:Guanidines; D006609:High Mobility Group Proteins / Q000097:blood*; D008239:Lysine / Q000097:blood*; D008297:Male; D011487:Protein Conformation; D013552:Swine; D014452:Ubiquitins / Q000097:blood*; D014508:Urea
chemical_list: D006146:Guanidines; D006609:High Mobility Group Proteins; D014452:Ubiquitins; D014508:Urea; D019791:Guanidine; D008239:Lysine; D004229:Dithiothreitol