ArticleFrontiers in plant science2015
Monitoring protein phosphorylation by acrylamide pendant Phos-Tag™ in various plants.
Article in Frontiers in plant science, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 14 papers.
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Who cites it
14 citing papers in PubMed, 16 citations in OpenAlex.
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- Advanced microscopy resolves dynamic localization patterns of stress-induced mitogen-activated protein kinase (SIMK) during alfalfa root hair interactions with Ensifer meliloti.Journal of experimental botany · 2023Article
- Overexpression of alfalfa SIMK promotes root hair growth, nodule clustering and shoot biomass production.Plant biotechnology journal · 2021Article
- Protoplast: A Valuable Toolbox to Investigate Plant Stress Perception and Response.Frontiers in plant science · 2021Review
- Structure of Arabidopsis HISTONE DEACETYLASE15.Plant physiology · 2020Article
- Biotechnological Perspectives of Omics and Genetic Engineering Methods in Alfalfa.Frontiers in plant science · 2020Review
- RNA stability protein ILF3 mediates cytokine-induced angiogenesis.FASEB journal : official publication of the Federation of American Societies for Experimental Biology · 2019Article
- The cotton MAPK kinase GhMPK20 negatively regulates resistance to Fusarium oxysporum by mediating the MKK4-MPK20-WRKY40 cascade.Molecular plant pathology · 2018Article
- BMI1, a new target of CK2α.Molecular cancer · 2017Article
- RhMKK9, a rose MAP KINASE KINASE gene, is involved in rehydration-triggered ethylene production in rose gynoecia.BMC plant biology · 2017Article
- Alfalfa Root Growth Rate Correlates with Progression of Microtubules during Mitosis and Cytokinesis as Revealed by Environmental Light-Sheet Microscopy.Frontiers in plant science · 2017Article
- Ser/Thr Phosphorylation Regulates the Fatty Acyl-AMP Ligase Activity of FadD32, an Essential Enzyme in Mycolic Acid Biosynthesis.The Journal of biological chemistry · 2016Article
- Phosphorylation of the Light-Harvesting Complex II Isoform Lhcb2 Is Central to State Transitions.Plant physiology · 2015Article
Corrections and comments
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Authors and funding
9 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The aim of the present study is to rationalize acrylamide pendant Phos-Tag™ in-gel discrimination of phosphorylated and non-phosphorylated plant protein species with standard immunoblot analysis, and optimize sample preparation, efficient electrophoretic separation and transfer. We tested variants of the method including extraction buffers suitable for preservation of phosphorylated protein species in crude extracts from plants and we addressed the importance of the cation (Mn(2+) or Zn(2+)) used in the gel recipe for efficient transfer to PVDF membranes for further immunoblot analysis. We demonstrate the monitoring of Medicago sativa stress-induced mitogen activated protein kinase (SIMK) in stress-treated wild type plants and transgenic SIMKK RNAi line. We further show the hyperosmotically-induced phosphorylation of the previously uncharacterized HvMPK4 of barley. The method is validated using inducible phosphorylation of barley and wheat α-tubulin and of Arabidopsis MPK6. Acrylamide pendant Phos-Tag™offers a flexible tool for studying protein phosphorylation in crops and Arabidopsis circumventing radioactive labeling and the use of phosphorylation specific antibodies.
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