PURIFICATION OF NITRATE REDUCTASE FROM SPINACH (SPINACEA-OLERACEA L) BY IMMUNOAFFINITY CHROMATOGRAPHY USING A MONOCLONAL-ANTIBODY

被引：25

作者：

FIDO, RJ

机构：

来源：

PLANT SCIENCE | 1987年 / 50卷 / 02期

关键词：

D O I：

10.1016/0168-9452(87)90146-4

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

NADH-Nitrate reductase (EC 1.6.6.1) from spinach (Spinacia oleracea L. v. Noorman) has been purified to apparent homogeneity by immunoaffinity chromatography using a monoclonal antibody linked covalently to Sepharose 4B followed by affinity chromatography. A pre-column of covalently linked non-immune rat .gamma. globulin prevented non-specific binding. The enzyme, released with 1 M KNO3, was purified 1550-fold to a specific activity of 24.8 .mu.mol NO2- produced min-1, mg protein-1 with a recovery of 60% of applied NADH-NR activity. Proteolytically ''nicked'' subunits, detected by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) were removed by 5''-AMP Sepharose chromatography (Fido and Notton, Plant Sci. Lett., 37 (1984) 87).

引用

页码：111 / 115

页数：5

共 21 条

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