Mapping of Plasma Membrane Proteins Interacting With Arabidopsis thaliana Flotillin 2

0492287 - ÚEB 2019 RIV CH eng J - Journal Article
Junková, P. - Daněk, Michal - Kocourková, Daniela - Brouzdová, Jitka - Kroumanová, Kristýna - Zelazny, E. - Janda, Martin - Hynek, R. - Martinec, Jan - Valentová, O.
Mapping of Plasma Membrane Proteins Interacting With Arabidopsis thaliana Flotillin 2.
Frontiers in Plant Science. Roč. 9, JUL 12 (2018), č. článku 991. ISSN 1664-462X. E-ISSN 1664-462X
R&D Projects: GA ČR GA14-09685S; GA MŠMT(CZ) 7AMB17FR005
Institutional support: RVO:61389030
Keywords : hypersensitive-induced reaction * independent endocytic pathway * syringae pv. tomato * mass-spectrometry * affinity-purification * lipid rafts * cell-death * h+-atpase * in-vivo * signal-transduction * Arabidopsis flotillin 2 * protein-protein interactions * immunopurification * mass spectrometry * splitubiquitin yeast system * plant-pathogen interaction * water transport * intracellular trafficking
OECD category: Cell biology
Impact factor: 4.106, year: 2018

Arabidopsis flotillin 2 (At5g25260) belongs to the group of plant flotillins, which are not well characterized. In contrast, metazoan flotillins are well known as plasma membrane proteins associated with membrane microdomains that act as a signaling hub. The similarity of plant and metazoan flotillins, whose functions most likely consist of affecting other proteins via protein-protein interactions, determines the necessity of detecting their interacting partners in plants. Nevertheless, identifying the proteins that form complexes on the plasma membrane is a challenging task due to their low abundance and hydrophobic character. Here we present an approach for mapping Arabidopsis thaliana flotillin 2 plasma membrane interactors, based on the immunoaffinity purification of crosslinked and enriched plasma membrane proteins with mass spectrometry detection. Using this approach, 61 proteins were enriched in the AtFlot-GFP plasma membrane fraction, and 19 of them were proposed to be flotillin 2 interaction partners. Among our proposed partners of Flot2, proteins playing a role in the plant response to various biotic and abiotic stresses were detected. Additionally, the use of the split-ubiquitin yeast system helped us to confirm that plasma-membrane ATPase 1, early-responsive to dehydration stress protein 4, syntaxin-71, harpin-induced protein-like 3, hypersensitive-induced response protein 2 and two aquaporin isoforms interact with flotillin 2 directly. Based on the results of our study and the reported properties of Flot2 interactors, we propose that Flot2 complexes may be involved in plant-pathogen interactions, water transport and intracellular trafficking.
Permanent Link: http://hdl.handle.net/11104/0285808