铁氧化还原蛋白-琼脂糖4B作为纯化铁氧化还原蛋白-NADP⁺还原酶的工具。

Ferredoxin-Sepharose 4B as a tool for the purification of ferredoxin-NADP+ reductase.

作者信息

Shin M, Oshino R

出版信息

J Biochem. 1978 Feb;83(2):357-61. doi: 10.1093/oxfordjournals.jbchem.a131921.

DOI:10.1093/oxfordjournals.jbchem.a131921

Abstract

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.

摘要

通过将溴化氰活化的琼脂糖4B与菠菜铁氧化还原蛋白反应，制备了固定在琼脂糖4B上的铁氧化还原蛋白。铁氧化还原蛋白-琼脂糖4B在稀缓冲溶液中结合铁氧化还原蛋白-NADP⁺还原酶（NADPH：铁氧化还原蛋白氧化还原酶，[EC 1.6.7.1]），并在高盐浓度下将其释放。通过将铁氧化还原蛋白-琼脂糖4B柱上的亲和吸附与常规纯化程序相结合，建立了一种制备该还原酶的新方法。使用新方法发现，菠菜中存在两种形式的铁氧化还原蛋白-NADP⁺还原酶，即FNR I和FNR II。对这两种成分的比较研究表明，FNR I可能是FNR II的二聚体。