Assaying the Effect of Peptide Treatment on H + -Pumping Activity in Plasma Membranes from Arabidopsis Seedlings.
| Autor: | Havshøi NW; Department of Plant and Environmental Sciences, Section for Transport Biology, University of Copenhagen, Frederiksberg, Denmark., Fuglsang AT; Department of Plant and Environmental Sciences, Section for Transport Biology, University of Copenhagen, Frederiksberg, Denmark. atf@plen.ku.dk. |
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| Jazyk: | angličtina |
| Zdroj: | Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2024; Vol. 2731, pp. 91-103. |
| DOI: | 10.1007/978-1-0716-3511-7_7 |
| Abstrakt: | Extracellular acidification or alkalization is a common response to many plant-signaling peptides and microbial elicitors. This may be a result of peptide-mediated regulation of plasma membrane-localized ion transporters, such as the plasma membrane H + -ATPase. Early responses to some signaling peptides can therefore be analyzed by assaying H + -pumping across the plasma membrane.We describe a set-up suited for the purification of plasma membranes by aqueous two-phase partitioning from a small sample of Arabidopsis seedlings. Seedlings are grown in a liquid culture, suited for the analysis of in vivo peptide treatment. Additionally, we describe how to measure the H + -pumping activity of the plasma membrane H + -ATPase using the fluorescent probe ACMA. (© 2024. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.) |
| Databáze: | MEDLINE |
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