Assaying the Effect of Peptide Treatment on H+-Pumping Activity in Plasma Membranes from Arabidopsis Seedlings

Nanna Weise Havshøi; Anja Thoe Fuglsang

doi:10.1007/978-1-0716-3511-7_7

Assaying the Effect of Peptide Treatment on H⁺-Pumping Activity in Plasma Membranes from Arabidopsis Seedlings

Nanna Weise Havshøi, Anja Thoe Fuglsang^*

^*Corresponding author for this work

Section for Transport Biology

Research output: Chapter in Book/Report/Conference proceeding › Book chapter › Research › peer-review

Abstract

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.

Original language	English
Title of host publication	Plant Peptide Hormones and Growth Factors
Number of pages	13
Publisher	Humana Press
Publication date	2024
Pages	91-103
Chapter	2
ISBN (Print)	978-1-0716-3510-0
ISBN (Electronic)	978-1-0716-3511-7
DOIs	https://doi.org/10.1007/978-1-0716-3511-7_7
Publication status	Published - 2024

Series	Methods in Molecular Biology
Volume	2731
ISSN	1064-3745

Bibliographical note

Publisher Copyright:
© 2024, The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.

Keywords

H pumping assay
H-ATPase
In vivo peptide treatment
Liquid seedling culture
Membrane vesicles
Plasma membrane
Two-phase partitioning

Access to Document

10.1007/978-1-0716-3511-7_7

Cite this

Assaying the Effect of Peptide Treatment on H⁺-Pumping Activity in Plasma Membranes from Arabidopsis Seedlings. / Havshøi, Nanna Weise ; Fuglsang, Anja Thoe.

Plant Peptide Hormones and Growth Factors. Humana Press, 2024. p. 91-103 (Methods in Molecular Biology, Vol. 2731).

Research output: Chapter in Book/Report/Conference proceeding › Book chapter › Research › peer-review

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