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https://hdl.handle.net/2440/55800
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Type: | Journal article |
Title: | Interference with the citrulline-based nitric oxide synthase assay by argininosuccinate lyase activity in Arabidopsis extracts |
Author: | Tischner, R. Galli, M. Heimer, Y. Bielefeld, S. Okamoto, M. Mack, A. Crawford, N. |
Citation: | The Federation of European Biochemical Societies (FEBS) Journal, 2007; 274(16):4238-4245 |
Publisher: | Blackwell Publishing Ltd |
Issue Date: | 2007 |
ISSN: | 1742-464X 1742-4658 |
Statement of Responsibility: | Rudolf Tischner, Mary Galli, Yair M. Heimer, Sarah Bielefeld, Mamoru Okamoto, Alyson Mack and Nigel M. Crawford |
Abstract: | There are many reports of an arginine-dependent nitric oxide synthase activity in plants; however, the gene(s) or protein(s) responsible for this activity have yet to be convincingly identified. To measure nitric oxide synthase activity, many studies have relied on a citrulline-based assay that measures the formation of L-citrulline from L-arginine using ion exchange chromatography. In this article, we report that when such assays are used with protein extracts from Arabidopsis, an arginine-dependent activity was observed, but it produced a product other than citrulline. TLC analysis identified the product as argininosuccinate. The reaction was stimulated by fumarate (> 500 microM), implicating the urea cycle enzyme argininosuccinate lyase (EC 4.3.2.1), which reversibly converts arginine and fumarate to argininosuccinate. These results indicate that caution is needed when using standard citrulline-based assays to measure nitric oxide synthase activity in plant extracts, and highlight the importance of verifying the identity of the product as citrulline. |
Keywords: | Arabidopsis argininosuccinate lyase citrulline nitric oxide |
DOI: | 10.1111/j.1742-4658.2007.05950.x |
Published version: | http://dx.doi.org/10.1111/j.1742-4658.2007.05950.x |
Appears in Collections: | Agriculture, Food and Wine publications Aurora harvest |
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