Higher Plant Nitrate Reductase: Structure, Mechanism and Modification

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Nitrate reductase (NR) is the enzyme that reduces soil nitrate to nitrite, a key step in the utilization of inorganic nitrogen fertilizer by plants. This proposal is aimed at elucidating the structure and mechanism of action of NR, which has been purified to homogeneity using affinity chromatography and monoclonal antibody columns. The NR gene from squash has been cloned. Efforts will be made to express this gene in a suitable host so that NR can be obtained in quantity for crystallization studies. The enzyme will be subjected to limited proteolysis and the fragments sequenced in order to identify the active sites. The results will enhance our understanding of how NR works, and may eventually enable plants to utilize nitrogen fertilizer more efficiently.