Dokument: Evidence for rapid hydrolysis of shoot‐derived sucrose using an ultrasensitive ratiometric matryoshka‐type MGlucoMeter sensor
| Titel: | Evidence for rapid hydrolysis of shoot‐derived sucrose using an ultrasensitive ratiometric matryoshka‐type MGlucoMeter sensor | |||||||
| URL für Lesezeichen: | https://docserv.uni-duesseldorf.de/servlets/DocumentServlet?id=72686 | |||||||
| URN (NBN): | urn:nbn:de:hbz:061-20260324-120348-0 | |||||||
| Kollektion: | Publikationen | |||||||
| Sprache: | Englisch | |||||||
| Dokumententyp: | Wissenschaftliche Texte » Artikel, Aufsatz | |||||||
| Medientyp: | Text | |||||||
| Autoren: | Ishikawa, Yuuma [Autor] Zöllner, Nora [Autor] Paradies, Susanne [Autor] Frommer, Wolf B. [Autor] | |||||||
| Dateien: |
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| Stichwörter: | technical advance | |||||||
| Beschreibung: | To enable sensitive in vivo monitoring of the glucose transport and metabolism, we developed a series of ultrasensitive and ratiometric genetically encoded sensors (MGlucoMeter) by inserting a Matryoshka dual fluorophore cassette consisting of cpsfGFP (circularly permuted superfolder GFP) and LSSmApple (Large Stokes Shift mApple) into the glucose-binding protein ttGBP (Thermus thermophilus glucose-binding protein) from Thermus thermophilus. The initial MGlucoMeter1.0 version was subjected to an alanine scan of the hinge region producing the more sensitive MGlucoMeter2.6 with a glucose-induced ΔF/F 0 change of 3.0, an affinity for glucose of 15 μM, and an approximate detection range of 1–215 μM. To generate variants suitable for in vivo measurements, a series of affinity mutants was generated by mutating two histidines predicted to be involved in substrate binding. MGlucoMeter2.6-353n (affinity 353 nM), MGlucoMeter2.6-15 μ
(affinity 15 μM ), MGlucoMeter2.6-700 μ (affinity 700 μM), MGlucoMeter2.6-1 m (affinity 1 m M), and MGlucoMeter2.6-7 m (affinity 7 m M) cover a combined detection range between ∼40 nM–55 mM . When expressed from a ubiquitous promoter in the cytosol of the Arabidopsis gene-silencing mutant rdr6 (RNA-dependent RNA polymerase 6), MGlucoMeter2.6-1 m reports time- and concentration-dependent accumulation of glucose in seedling roots after external addition of glucose. The sensor also detected rapid release of sugars in the root tip and rapid hydrolysis of the shoot-derived sucrose. | |||||||
| Rechtliche Vermerke: | Originalveröffentlichung:
Ishikawa, Y., Zöllner, N., Paradies, S., & Frommer, W. B. (2026). Evidence for rapid hydrolysis of shoot‐derived sucrose using an ultrasensitive ratiometric matryoshka‐type MGlucoMeter sensor. The plant journal, 125(5), Article e70778. https://doi.org/10.1111/tpj.70778 | |||||||
| Lizenz: |  Dieses Werk ist lizenziert unter einer Creative Commons Namensnennung 4.0 International Lizenz | |||||||
| Fachbereich / Einrichtung: | Mathematisch- Naturwissenschaftliche Fakultät | |||||||
| Dokument erstellt am: | 24.03.2026 | |||||||
| Dateien geändert am: | 24.03.2026 |
