Please use this identifier to cite or link to this item: http://hdl.handle.net/1822/51844

TitleAnalytical and Fluorimetric Methods for the Characterization of the Transmembrane Transport of Specialized Metabolites in Plants
Author(s)Carqueijeiro, Inês
Martins, Viviana Maria Varajão
Noronha, Henrique Luis Silva de
Gerós, H.
Sottomayor, Mariana
KeywordsABC transporters
ACMA
Alkaloids
BY-2 cells
Catharanthus roseus
H+-antiport
MATE transporters
Tobacco
Tonoplast vesicles
Transmembrane transport
Transmembrane pH gradient
Uptake assay
Issue dateFeb-2016
PublisherHumana Press
CitationCarqueijeiro I., Martins V., Noronha H., Gerós H., Sottomayor M. (2016) Analytical and Fluorimetric Methods for the Characterization of the Transmembrane Transport of Specialized Metabolites in Plants. In: Fett-Neto A. (eds) Biotechnology of Plant Secondary Metabolism. Methods in Molecular Biology, vol 1405. Humana Press, New York, NY
Abstract(s)The characterization of membrane transport of specialized metabolites is essential to understand their metabolic fluxes and to implement metabolic engineering strategies towards the production of increased levels of these valuable metabolites. Here, we describe a set of procedures to isolate tonoplast membranes, to check their purity and functionality, and to characterize their transport properties. Transport is assayed directly by HPLC analysis and quantification of the metabolites actively accumulated in the vesicles, and indirectly using the pH sensitive fluorescent probe ACMA (9-amino-6- chloro-2-methoxyacridine), when a proton antiport is involved.
TypebookPart
URIhttp://hdl.handle.net/1822/51844
ISBN978-1-4939-3391-4
e-ISBN978-1-4939-3393-8
DOI10.1007/978-1-4939-3393-8_12
Publisher versionhttps://link.springer.com/protocol/10.1007/978-1-4939-3393-8_12#citeas
AccessclosedAccess
Appears in Collections:DBio - Livros e Capítulos de Livros/Books and Books Chapters

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