Ferredoxin-Sepharose 4B as a tool for the purification of ferredoxin-NADP+ reductase.
Ferredoxin immobilized on Sepharose 4B was prepared by reaction of CNBr-Sepharose 4B with spinach ferredoxin. The ferredoxin-Sepharose 4B conjugated ferredoxin-NADP+ reductase (NADPH: ferredoxin oxidoreductase, [EC 1.6.7.1]) in dilute buffer solution and released it in high salt concentrations. A novel method of preparation for the reductase was established by a combination of affinity adsorption on the ferredoxin-Sepharose 4B column with usual purification procedures. It was found using the new method, that there are two forms of ferredoxin-NADP+ reductase, FNR I and FNR II, in spinach. Comparative studies of the two components suggest that FNR I may be a dimer of FNR II.
D002846:Chromatography, Affinity; D005287:Ferredoxin-NADP Reductase / Q000302:isolation & purification*; D005288:Ferredoxins; D046911:Macromolecular Substances; D009247:NADH, NADPH Oxidoreductases / Q000302:isolation & purification*; D010944:Plants / Q000201:enzymology; D011134:Polysaccharides; D012685:Sepharose
D005288:Ferredoxins; D046911:Macromolecular Substances; D011134:Polysaccharides; D012685:Sepharose; D005287:Ferredoxin-NADP Reductase; D009247:NADH, NADPH Oxidoreductases