title: SS-interchanged and oxidized isomers of bovine serum albumin separated by isoelectric focusing.
abstract: Column isoelectric focusing separates commercial bovine serum albumin in 5 fractions with isoionic points in the vicinity of that of mercaptalbumin (pI 5.24). About 20% of the bovine albumin have isoionic points higher than mercaptalbumin and are split into two fractions, both recognized as SS-interchanges isomers: (1) pI 5.39 is the "aged" albumin described by Nikkel and Foster (1971, Biochemistry 10, 4479); (2) pI 5.45 represents a further degree of SS-interchange, catalyzed by small amounts of cysteine in the solution ('cysteine-aged' albumin). In 6 M urea the "cysteine-aged" albumin is electrofocused to the same pH value as mercaptalbumin. In 6 M urea 40% of commercial albumin focuses in 3 fractions with isoionic points lower than mercaptalbumin. This percentage will increase during incubation at oxidizing conditions ("oxidized" albumin). Electrofocused in water the oxidized fractions have isoionic points at pI 5.28, 5.18 and 5.12, respectively. The shifts in isoionic point of the "oxidized" albumins are caused by irreversible changes in the primary structure. Although the free SH group of albumin is oxidized during the oxidation reaction, the observed changes in isoionic points are caused by modifications of some other amino acid residues. Both "cysteine-ageing" and "oxidation" are inhibited by alkylation of the SH group. "Cysteine-ageing" is furthermore inhibited when the bovine albumin is "oxidized".
mesh_terms: D000818:Animals; D001665:Binding Sites; D002417:Cattle; D004220:Disulfides; D006146:Guanidines; D007460:Iodoacetamide; D007525:Isoelectric Focusing; D007536:Isomerism; D008623:Mercaptoethanol; D010084:Oxidation-Reduction; D011485:Protein Binding; D011487:Protein Conformation; D012710:Serum Albumin, Bovine; D013056:Spectrophotometry, Ultraviolet; D014508:Urea
chemical_list: D004220:Disulfides; D006146:Guanidines; D012710:Serum Albumin, Bovine; D008623:Mercaptoethanol; D014508:Urea; D007460:Iodoacetamide