Wallsgrove R M, Lea P J, Miflin B J
Biochemistry Department, Rothamsted Experimental Station, Harpenden, Herts, AL5 2JQ, United Kingdom.
Plant Physiol. 1979 Feb;63(2):232-6. doi: 10.1104/pp.63.2.232.
Protoplasts obtained from expanded leaves of Pisum sativum have been used for the isolation of cell organelles and the subsequent study of the intracellular distribution of the enzymes of nitrate assimilation. The protoplasts were ruptured in a suitable medium and the total lysate subjected to sucrose density gradient centrifugation. Of the total chlorophyll more than 80% was recovered in intact chloroplasts. Nitrite reductase and glutamate synthase were found to be located wholly in the chloroplast. Glutamine synthetase was distributed between the chloroplast and the cytoplasm, with a maximum of 60% of the former. A possible role of the cytoplasmic enzyme is discussed in relation to photorespiration. There was no evidence for the association of nitrate reductase with any cell organelle or membrane fraction.
从豌豆展开叶中获得的原生质体已被用于分离细胞器，并随后研究硝酸盐同化酶在细胞内的分布。原生质体在合适的培养基中破裂，总裂解物进行蔗糖密度梯度离心。完整叶绿体中回收了超过80%的总叶绿素。发现亚硝酸还原酶和谷氨酸合酶完全位于叶绿体中。谷氨酰胺合成酶分布在叶绿体和细胞质之间，前者最多占60%。讨论了细胞质酶与光呼吸相关的可能作用。没有证据表明硝酸还原酶与任何细胞器或膜组分有关联。
