| DAF-fluorescence without NO: Elicitor treated tobacco cells produce fluorescing DAF-derivatives not related to DAF-2 triazol. | |
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MedLine Citation:
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PMID: 22683597 Owner: NLM Status: Publisher |
Abstract/OtherAbstract:
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Diaminofluorescein-dyes (DAFs) are widely used for visualizing NO(•) production in biological systems. Here it was examined whether DAF-fluorescence could be evoked by other means than nitrosation. Tobacco (Nicotiana tabacum) suspension cells treated with the fungal elicitor cryptogein released compound(s) which gave a fluorescence increase in the cell-free filtrate after addition of DAF-2 or DAF-FM or DAR-4M. DAF-reactive compounds were relatively stable and identified as reaction products of H(2)O(2) plus apoplastic peroxidase (PO). CPTIO prevented formation of these products. Horseradish-peroxidase (HR-PO) plus H(2)O(2) also generated DAF-fluorescence in vitro. Using RP-HPLC with fluorescence detection, DAF derivatives were further analyzed. In filtrates from cryptogein-treated cells, fluorescence originated from two novel DAF-derivatives also obtained in vitro with DAF-2+HR-PO+H(2)O(2). DAF-2T was only detected when an NO donor (DEA-NO) was present. Using high resolution mass spectrometry, the two above-described novel DAF-reaction products were tentatively identified as dimers. In cells preloaded with DAF-2 DA and incubated with or without cryptogein, DAF-fluorescence originated from a complex pattern of multiple products different from those obtained in vitro. One specific peak was responsive to exogenous H(2)O(2), and another, minor peak eluted at or close to DAF-2T. Thus, in contrast to the prevailing opinion, DAF-2 can be enzymatically converted into a variety of highly fluorescing derivatives, both inside and outside cells, of which none (outside) or only a minor part (inside) appeared NO(•) dependent. Accordingly, DAF-fluorescence and its prevention by cPTIO do not necessarily indicate NO(•) production. |
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Authors:
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Stefan Rümer; Markus Krischke; Agnes Fekete; Martin J Mueller; Werner M Kaiser |
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Publication Detail:
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Type: JOURNAL ARTICLE Date: 2012-6-5 |
Journal Detail:
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Title: Nitric oxide : biology and chemistry / official journal of the Nitric Oxide Society Volume: - ISSN: 1089-8611 ISO Abbreviation: - Publication Date: 2012 Jun |
Date Detail:
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Created Date: 2012-6-11 Completed Date: - Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 9709307 Medline TA: Nitric Oxide Country: - |
Other Details:
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Languages: ENG Pagination: - Citation Subset: - |
Copyright Information:
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Copyright © 2012. Published by Elsevier Inc. |
Affiliation:
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University of Wuerzburg, Julius-von-Sachs Institute of Biosciences, Julius-von-Sachs-Platz 2, D-97082 Wuerzburg, Germany. |
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From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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