Artificial Fluorescent Glucosinolates (F-GSLs) Are Transported by the Glucosinolate Transporters GTR1/2/3

Christa Kanstrup, Claire C. Jimidar, Josip Tomas, Giuliano Cutolo, Christoph Crocoll, Marie Schuler, Philipp Klahn, Arnaud Tatibouët, Hussam Hassan Nour-Eldin*

*Corresponding author af dette arbejde

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Abstract

The glucosinolate transporters 1/2/3 (GTR1/2/3) from the Nitrate and Peptide transporter Family (NPF) play an essential role in the transport, accumulation, and distribution of the specialized plant metabolite glucosinolates. Due to representing both antinutritional and health-promoting compounds, there is increasing interest in characterizing GTRs from various plant species. We generated seven artificial glucosinolates (either aliphatic or benzenic) bearing different fluorophores (Fluorescein, BODIPY, Rhodamine, Dansylamide, and NBD) and investigated the ability of GTR1/2/3 from Arabidopsis thaliana to import the fluorescent glucosinolates (F-GSLs) into oocytes from Xenopus laevis. Five out of the seven F-GSLs synthesized were imported by at least one of the GTRs. GTR1 and GTR2 were able to import three F-GSLs actively above external concentration, while GTR3 imported only one actively. Competition assays indicate that the F-GSLs are transported by the same mechanism as non-tagged natural glucosinolates. The GTR-mediated F-GSL uptake is detected via a rapid and sensitive assay only requiring simple fluorescence measurements on a standard plate reader. This is highly useful in investigations of glucosinolate transport function and provides a critical prerequisite for elucidating the relationship between structure and function through high-throughput screening of GTR mutant libraries. The F-GSL themselves may also be suitable for future studies on glucosinolate transport in vivo.

OriginalsprogEngelsk
Artikelnummer920
TidsskriftInternational Journal of Molecular Sciences
Vol/bind24
Udgave nummer2
Antal sider26
ISSN1661-6596
DOI
StatusUdgivet - 2023

Bibliografisk note

Funding Information:
The design and synthesis of certain F-GSLs in this work were carried out within the framework of the SMART BIOTECS alliance between the Technische Universität Braunschweig and the Leibniz Universität Hannover (P.K.). This initiative is supported by the Ministry of Science and Culture (MWK) of Lower Saxony, Germany. Financial support by the Max-Buchner Research Foundation (Max-Buchner Research Fellowship, P.K.) is gratefully acknowledged. Part of the F-GSLs was designed and synthesized with the financial support of projects CHemBio (FEDER-FSE 2014-2020-EX003677), Techsab (FEDER-FSE 2014-2020-EX011313), QUALICHIM (APR-IA-PF 2021-00149467), the RTR Motivhealth (2019-00131403) and the Labex programs SYNORG (ANR-11-LABX-0029) and IRON (ANR-11-LABX-0018-01), and the SALSA platform is gratefully acknowledged for analytical support. The transport assay experimental work was supported by the Danish Research Foundation (Grant no. DNRF99) and the Human Frontier Science Program (RGY0075/2020). The content of this work is solely the responsibility of the authors and does not necessarily represent the official views of the funding agencies.

Publisher Copyright:
© 2023 by the authors.

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