title: Purification by hydrophobic chromatography of soluble cytochrome b5 of human erythrocytes.
abstract: Soluble cytochrome b5 of human erythrocytes was purified very effectively by hydrophobic chromatography using a butyl-Toyopearl 650 column. Cytochrome b5 was adsorbed tightly on the column in the presence of 60% saturated ammonium sulfate, and was eluted at 40% saturation of ammonium sulfate in the elution buffer. The chromatography gave a good yield of cytochrome b5 of the highest purity so far reported as estimated from the 414 nm to 280 nm absorbance ratio of the oxidized form of the cytochrome b5. The value obtained with the cytochrome b5 purified in this study was 6.57, and is higher than the previously reported highest value of 6.4 (Hultquist, D.E., Dean, R.T. and Douglas, R.H. (1974) Biochem. Biophys. Res. Commun. 60, 28-34). Spectral properties including molecular absorption coefficients were determined using the cytochrome b5 purified by this method.
authors.affiliation: Department of Biochemistry, Medical College of Oita, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Yubisui
authors.forename: K
authors.initials: K
authors.lastname: Murakami
authors.forename: M
authors.initials: M
authors.lastname: Takeshita
authors.forename: T
authors.initials: T
authors.lastname: Takano
mesh_terms: D000596:Amino Acids / Q000032:analysis; D000645:Ammonium Sulfate; D000818:Animals; D002417:Cattle; D002645:Chickens; D002845:Chromatography; D002848:Chromatography, DEAE-Cellulose; D002850:Chromatography, Gel; D003573:Cytochrome b Group / Q000097:blood*; D015786:Cytochromes b5; D004912:Erythrocytes / Q000032:analysis*; D005260:Female; D006801:Humans; D008297:Male; D008862:Microsomes, Liver / Q000032:analysis; D008970:Molecular Weight; D013053:Spectrophotometry
chemical_list: D000596:Amino Acids; D003573:Cytochrome b Group; D015786:Cytochromes b5; D000645:Ammonium Sulfate