The Impact of Nitrile-Specifier Proteins on Indolic Carbinol and Nitrile Formation in Homogenates of <i>Arabidopsis thaliana</i>
Glucosinolates, specialized metabolites of the Brassicales including <i>Brassica</i> crops and <i>Arabidopsis thaliana</i>, have attracted considerable interest as chemical defenses and health-promoting compounds. Their biological activities are mostly due to breakdown produc...
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| Format: | Article |
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MDPI AG
2022-11-01
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| Series: | Molecules |
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| Online Access: | https://www.mdpi.com/1420-3049/27/22/8042 |
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| author | Eleanor C. M. Chroston Annika Hielscher Matthias Strieker Ute Wittstock |
| author_facet | Eleanor C. M. Chroston Annika Hielscher Matthias Strieker Ute Wittstock |
| author_sort | Eleanor C. M. Chroston |
| collection | DOAJ |
| description | Glucosinolates, specialized metabolites of the Brassicales including <i>Brassica</i> crops and <i>Arabidopsis thaliana</i>, have attracted considerable interest as chemical defenses and health-promoting compounds. Their biological activities are mostly due to breakdown products formed upon mixing with co-occurring myrosinases and specifier proteins, which can result in multiple products with differing properties, even from a single glucosinolate. Whereas product profiles of aliphatic glucosinolates have frequently been reported, indole glucosinolate breakdown may result in complex mixtures, the analysis of which challenging. The aim of this study was to assess the breakdown of indole glucosinolates in <i>A. thaliana</i> root and rosette homogenates and to test the impact of nitrile-specifier proteins (NSPs) on product profiles. To develop a GC-MS-method for quantification of carbinols and nitriles derived from three prominent indole glucosinolates, we synthesized standards, established derivatization conditions, determined relative response factors and evaluated applicability of the method to plant homogenates. We show that carbinols are more dominant among the detected products in rosette than in root homogenates of wild-type and NSP1- or NSP3-deficient mutants. NSP1 is solely responsible for nitrile formation in rosette homogenates and is the major NSP for indolic nitrile formation in root homogenates, with no contribution from NSP3. These results will contribute to the understanding of the roles of NSPs in plants. |
| first_indexed | 2024-03-09T18:06:42Z |
| format | Article |
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| institution | Directory Open Access Journal |
| issn | 1420-3049 |
| language | English |
| last_indexed | 2024-03-09T18:06:42Z |
| publishDate | 2022-11-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Molecules |
| spelling | doaj.art-28835830c55d49948c4e143662e69dcb2023-11-24T09:25:29ZengMDPI AGMolecules1420-30492022-11-012722804210.3390/molecules27228042The Impact of Nitrile-Specifier Proteins on Indolic Carbinol and Nitrile Formation in Homogenates of <i>Arabidopsis thaliana</i>Eleanor C. M. Chroston0Annika Hielscher1Matthias Strieker2Ute Wittstock3Institute of Pharmaceutical Biology, Technische Universität Braunschweig, Mendelssohnstr. 1, D-38106 Braunschweig, GermanyInstitute of Pharmaceutical Biology, Technische Universität Braunschweig, Mendelssohnstr. 1, D-38106 Braunschweig, GermanyInstitute of Pharmaceutical Biology, Technische Universität Braunschweig, Mendelssohnstr. 1, D-38106 Braunschweig, GermanyInstitute of Pharmaceutical Biology, Technische Universität Braunschweig, Mendelssohnstr. 1, D-38106 Braunschweig, GermanyGlucosinolates, specialized metabolites of the Brassicales including <i>Brassica</i> crops and <i>Arabidopsis thaliana</i>, have attracted considerable interest as chemical defenses and health-promoting compounds. Their biological activities are mostly due to breakdown products formed upon mixing with co-occurring myrosinases and specifier proteins, which can result in multiple products with differing properties, even from a single glucosinolate. Whereas product profiles of aliphatic glucosinolates have frequently been reported, indole glucosinolate breakdown may result in complex mixtures, the analysis of which challenging. The aim of this study was to assess the breakdown of indole glucosinolates in <i>A. thaliana</i> root and rosette homogenates and to test the impact of nitrile-specifier proteins (NSPs) on product profiles. To develop a GC-MS-method for quantification of carbinols and nitriles derived from three prominent indole glucosinolates, we synthesized standards, established derivatization conditions, determined relative response factors and evaluated applicability of the method to plant homogenates. We show that carbinols are more dominant among the detected products in rosette than in root homogenates of wild-type and NSP1- or NSP3-deficient mutants. NSP1 is solely responsible for nitrile formation in rosette homogenates and is the major NSP for indolic nitrile formation in root homogenates, with no contribution from NSP3. These results will contribute to the understanding of the roles of NSPs in plants.https://www.mdpi.com/1420-3049/27/22/8042indole glucosinolatesindolic carbinolindolic nitrilederivatizationGC-MSquantification |
| spellingShingle | Eleanor C. M. Chroston Annika Hielscher Matthias Strieker Ute Wittstock The Impact of Nitrile-Specifier Proteins on Indolic Carbinol and Nitrile Formation in Homogenates of <i>Arabidopsis thaliana</i> Molecules indole glucosinolates indolic carbinol indolic nitrile derivatization GC-MS quantification |
| title | The Impact of Nitrile-Specifier Proteins on Indolic Carbinol and Nitrile Formation in Homogenates of <i>Arabidopsis thaliana</i> |
| title_full | The Impact of Nitrile-Specifier Proteins on Indolic Carbinol and Nitrile Formation in Homogenates of <i>Arabidopsis thaliana</i> |
| title_fullStr | The Impact of Nitrile-Specifier Proteins on Indolic Carbinol and Nitrile Formation in Homogenates of <i>Arabidopsis thaliana</i> |
| title_full_unstemmed | The Impact of Nitrile-Specifier Proteins on Indolic Carbinol and Nitrile Formation in Homogenates of <i>Arabidopsis thaliana</i> |
| title_short | The Impact of Nitrile-Specifier Proteins on Indolic Carbinol and Nitrile Formation in Homogenates of <i>Arabidopsis thaliana</i> |
| title_sort | impact of nitrile specifier proteins on indolic carbinol and nitrile formation in homogenates of i arabidopsis thaliana i |
| topic | indole glucosinolates indolic carbinol indolic nitrile derivatization GC-MS quantification |
| url | https://www.mdpi.com/1420-3049/27/22/8042 |
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