DAF-fluorescence without NO: elicitor treated tobacco cells produce fluorescing DAF-derivatives not related to DAF-2 triazol.
Identifieur interne : 001562 ( Main/Exploration ); précédent : 001561; suivant : 001563DAF-fluorescence without NO: elicitor treated tobacco cells produce fluorescing DAF-derivatives not related to DAF-2 triazol.
Auteurs : Stefan Rümer [Allemagne] ; Markus Krischke ; Agnes Fekete ; Martin J. Mueller ; Werner M. KaiserSource :
- Nitric oxide : biology and chemistry [ 1089-8611 ] ; 2012.
Descripteurs français
- KwdFr :
- Espace intracellulaire (composition chimique), Espace intracellulaire (métabolisme), Espèces réactives de l'oxygène (métabolisme), Extrait cellulaire (composition chimique), Fluorescéine (composition chimique), Fluorescéine (métabolisme), Indicateurs et réactifs (composition chimique), Indicateurs et réactifs (métabolisme), Monoxyde d'azote (composition chimique), Monoxyde d'azote (métabolisme), Myeloperoxidase (composition chimique), Myeloperoxidase (métabolisme), Peroxyde d'hydrogène (composition chimique), Peroxyde d'hydrogène (métabolisme), Protéines fongiques (pharmacologie), Rhodamines (composition chimique), Rhodamines (métabolisme), Spectrométrie de fluorescence (MeSH), Tabac (composition chimique), Tabac (cytologie), Tabac (effets des médicaments et des substances chimiques), Tabac (métabolisme).
- MESH :
- composition chimique : Espace intracellulaire, Extrait cellulaire, Fluorescéine, Indicateurs et réactifs, Monoxyde d'azote, Myeloperoxidase, Peroxyde d'hydrogène, Rhodamines, Tabac.
- cytologie : Tabac.
- effets des médicaments et des substances chimiques : Tabac.
- métabolisme : Espace intracellulaire, Espèces réactives de l'oxygène, Fluorescéine, Indicateurs et réactifs, Monoxyde d'azote, Myeloperoxidase, Peroxyde d'hydrogène, Rhodamines, Tabac.
- pharmacologie : Protéines fongiques.
- Spectrométrie de fluorescence.
English descriptors
- KwdEn :
- Cell Extracts (chemistry), Fluorescein (chemistry), Fluorescein (metabolism), Fungal Proteins (pharmacology), Hydrogen Peroxide (chemistry), Hydrogen Peroxide (metabolism), Indicators and Reagents (chemistry), Indicators and Reagents (metabolism), Intracellular Space (chemistry), Intracellular Space (metabolism), Nitric Oxide (chemistry), Nitric Oxide (metabolism), Peroxidase (chemistry), Peroxidase (metabolism), Reactive Oxygen Species (metabolism), Rhodamines (chemistry), Rhodamines (metabolism), Spectrometry, Fluorescence (MeSH), Tobacco (chemistry), Tobacco (cytology), Tobacco (drug effects), Tobacco (metabolism).
- MESH :
- chemical , chemistry : Cell Extracts, Fluorescein, Hydrogen Peroxide, Indicators and Reagents, Nitric Oxide, Peroxidase, Rhodamines.
- chemical , metabolism : Fluorescein, Hydrogen Peroxide, Indicators and Reagents, Nitric Oxide, Peroxidase, Reactive Oxygen Species, Rhodamines.
- chemical , pharmacology : Fungal Proteins.
- chemistry : Intracellular Space, Tobacco.
- cytology : Tobacco.
- drug effects : Tobacco.
- metabolism : Intracellular Space, Tobacco.
- Spectrometry, Fluorescence.
Abstract
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.
DOI: 10.1016/j.niox.2012.05.007
PubMed: 22683597
Affiliations:
Links toward previous steps (curation, corpus...)
Le document en format XML
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<term>Fluorescein (metabolism)</term>
<term>Fungal Proteins (pharmacology)</term>
<term>Hydrogen Peroxide (chemistry)</term>
<term>Hydrogen Peroxide (metabolism)</term>
<term>Indicators and Reagents (chemistry)</term>
<term>Indicators and Reagents (metabolism)</term>
<term>Intracellular Space (chemistry)</term>
<term>Intracellular Space (metabolism)</term>
<term>Nitric Oxide (chemistry)</term>
<term>Nitric Oxide (metabolism)</term>
<term>Peroxidase (chemistry)</term>
<term>Peroxidase (metabolism)</term>
<term>Reactive Oxygen Species (metabolism)</term>
<term>Rhodamines (chemistry)</term>
<term>Rhodamines (metabolism)</term>
<term>Spectrometry, Fluorescence (MeSH)</term>
<term>Tobacco (chemistry)</term>
<term>Tobacco (cytology)</term>
<term>Tobacco (drug effects)</term>
<term>Tobacco (metabolism)</term>
</keywords>
<keywords scheme="KwdFr" xml:lang="fr"><term>Espace intracellulaire (composition chimique)</term>
<term>Espace intracellulaire (métabolisme)</term>
<term>Espèces réactives de l'oxygène (métabolisme)</term>
<term>Extrait cellulaire (composition chimique)</term>
<term>Fluorescéine (composition chimique)</term>
<term>Fluorescéine (métabolisme)</term>
<term>Indicateurs et réactifs (composition chimique)</term>
<term>Indicateurs et réactifs (métabolisme)</term>
<term>Monoxyde d'azote (composition chimique)</term>
<term>Monoxyde d'azote (métabolisme)</term>
<term>Myeloperoxidase (composition chimique)</term>
<term>Myeloperoxidase (métabolisme)</term>
<term>Peroxyde d'hydrogène (composition chimique)</term>
<term>Peroxyde d'hydrogène (métabolisme)</term>
<term>Protéines fongiques (pharmacologie)</term>
<term>Rhodamines (composition chimique)</term>
<term>Rhodamines (métabolisme)</term>
<term>Spectrométrie de fluorescence (MeSH)</term>
<term>Tabac (composition chimique)</term>
<term>Tabac (cytologie)</term>
<term>Tabac (effets des médicaments et des substances chimiques)</term>
<term>Tabac (métabolisme)</term>
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<term>Fluorescein</term>
<term>Hydrogen Peroxide</term>
<term>Indicators and Reagents</term>
<term>Nitric Oxide</term>
<term>Peroxidase</term>
<term>Rhodamines</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Fluorescein</term>
<term>Hydrogen Peroxide</term>
<term>Indicators and Reagents</term>
<term>Nitric Oxide</term>
<term>Peroxidase</term>
<term>Reactive Oxygen Species</term>
<term>Rhodamines</term>
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<term>Tobacco</term>
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<term>Extrait cellulaire</term>
<term>Fluorescéine</term>
<term>Indicateurs et réactifs</term>
<term>Monoxyde d'azote</term>
<term>Myeloperoxidase</term>
<term>Peroxyde d'hydrogène</term>
<term>Rhodamines</term>
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<keywords scheme="MESH" qualifier="cytology" xml:lang="en"><term>Tobacco</term>
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<keywords scheme="MESH" qualifier="drug effects" xml:lang="en"><term>Tobacco</term>
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<keywords scheme="MESH" qualifier="effets des médicaments et des substances chimiques" xml:lang="fr"><term>Tabac</term>
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<keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Espace intracellulaire</term>
<term>Espèces réactives de l'oxygène</term>
<term>Fluorescéine</term>
<term>Indicateurs et réactifs</term>
<term>Monoxyde d'azote</term>
<term>Myeloperoxidase</term>
<term>Peroxyde d'hydrogène</term>
<term>Rhodamines</term>
<term>Tabac</term>
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<keywords scheme="MESH" qualifier="pharmacologie" xml:lang="fr"><term>Protéines fongiques</term>
</keywords>
<keywords scheme="MESH" xml:lang="en"><term>Spectrometry, Fluorescence</term>
</keywords>
<keywords scheme="MESH" xml:lang="fr"><term>Spectrométrie de fluorescence</term>
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<front><div type="abstract" xml:lang="en">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.</div>
</front>
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<Abstract><AbstractText>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.</AbstractText>
<CopyrightInformation>Copyright © 2012 Elsevier Inc. All rights reserved.</CopyrightInformation>
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<QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName>
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<ArticleId IdType="doi">10.1016/j.niox.2012.05.007</ArticleId>
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