title: Immobilized ferredoxins for affinity chromatography of ferredoxin-dependent enzymes.
abstract: An immobilized ferredoxin more stable than the conventional immobilized spinach ferrodoxin was prepared by reacting CNBr-Sepharose with ferredoxins isolated from barley and Synechococcus vulcanus, a thermophilic blue-green alga. The dissociation constants of immobilized ferredoxin from spinach, barley and S. vulcanus for spinach ferredoxin-NADP reductase were 0.922, 2.505 and 5.209 microM, respectively, whereas those for barley ferredoxin-NADP reductase were 1.159, 0.579 and 2.851 microM, respectively. The order of stability was S. vulcanus greater than barley greater than spinach. The immobilized ferredoxin was applied to the simultaneous detection of ferredoxin-dependent enzymes in spinach chloroplasts. Over 20 polypeptides were detected. Synechococcus ferredoxin could also be immobilized on a Toyopearl gel and repeatedly used in an automated high-performance liquid chromatographic system.
authors.affiliation: Department of Biology, Faculty of Science, Kobe University, Japan.
authors.forename: N
authors.initials: N
authors.lastname: Sakihama
authors.forename: K
authors.initials: K
authors.lastname: Nagai
authors.forename: H
authors.initials: H
authors.lastname: Ohmori
authors.forename: H
authors.initials: H
authors.lastname: Tomizawa
authors.forename: M
authors.initials: M
authors.lastname: Tsujita
authors.forename: M
authors.initials: M
authors.lastname: Shin
mesh_terms: D055598:Chemical Phenomena; D002627:Chemistry, Physical; D002736:Chloroplasts / Q000737:chemistry; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D005287:Ferredoxin-NADP Reductase / Q000302:isolation & purification*; D005288:Ferredoxins / Q000302:isolation & purification*; D005782:Gels; D010944:Plants / Q000737:chemistry; D012685:Sepharose
chemical_list: D005288:Ferredoxins; D005782:Gels; D012685:Sepharose; D005287:Ferredoxin-NADP Reductase