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Phosphoproteome profiling under low nitrate and high nitrate in Arabisopsis wild type roots

ABSTRACT: In this project, we treated Arabidopsis wild type (Col-0) with low nitrate (0.2 mM KNO3) and high nitrate (5 mM KNO3) after nitrate starvation. Next we harvested root, and then we performed memebrane fraction and cytosolic fraction isolation for trypsin digestion and phosphopeptide enrichment.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Arabidopsis Thaliana (mouse-ear Cress)

TISSUE(S): Plant Cell, Root

SUBMITTER: Xuna Wu

LAB HEAD: Waltraud Schulze

PROVIDER: PXD014146 | Pride | 2024-05-21

REPOSITORIES: Pride

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			Action	DRS
	MaxQuant_Output.zip	Other
	WT_200uMNO3_15_1.raw	Raw
	WT_200uMNO3_15_2.raw	Raw
	WT_200uMNO3_15_3.raw	Raw
	WT_200uMNO3_15_4.raw	Raw

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Publications

Phosphoregulation in the N-terminus of NRT2.1 affects nitrate uptake by controlling the interaction of NRT2.1 with NAR2.1 and kinase HPCAL1 in Arabidopsis.

Li Zhi Z Na Wu Xu X Jacquot Aurore A Chaput Valentin V Adamo Mattia M Neuhäuser Benjamin B Straub Tatsiana T Lejay Laurence L Schulze Waltraud X WX

Journal of experimental botany 20240301 7

NRT2.1, the major high affinity nitrate transporter in roots, can be phosphorylated at five different sites within the N- and the C-terminus. Here, we characterized the functional relationship of two N-terminal phosphorylation sites, S21 and S28, in Arabidopsis. Based on a site-specific correlation network, we identified a receptor kinase (HPCAL1, AT5G49770), phosphorylating NRT2.1 at S21 and resulting in active nitrate uptake. HPCAL1 itself was regulated by phosphorylation at S839 and S870 with ...[more]

PMID: 38066636

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Phosphoproteomics of WT and nrt1.1 under high nitrate and low nitrate

Project description:We performed a comparative phosphoproteomic analysis of Arabidopsis root membrane proteins of WT and nrt1.1-1 under low nitrate (LN) and high nitrate (HN) availability in order to identify the downstream components that may be involved in NRT1.1-dependent LR growth and signal transduction.

2024-02-22 | PXD048853 | Pride

search substrates of receptor kinase complex SIRK1/QSK1

Project description:In this project, we are searching phosphorylation targets of receptor kinases complexes SIRK1/QSK1, especially to find the phosphorylation status of aquaporin PIP2F in sirk single mutant and sirk/qsk1 double mutant.

2019-11-12 | PXD011284 | Pride

Day-night phosphoproteome analysis in WT, pgm mutant and sweet11/sweet12 mutant

Project description:In this progect, we anallyzed the phosphoproteome change of shoot and root from WT pgm mutant and sweet11/sweet12 mutant at the end of day and the end of night. The purpose is to understand the role of sugar distribution in shoot and root phopshoproteome.

2022-08-12 | PXD031942 | Pride

Nitrate Starvation time course membrane protein phosphoproteomics

Project description:Nitrate nutrtition was withdrawn from Arabiodpsis plants to study early responses upon nitrate depletion. Plants were grown under full nutrition (3mM nitrate) for three weeks and then transferred to nutrient solution without nitrate for 15 minutes and 3 hours.

2024-05-21 | PXD015390 | Pride

sucrose-induced interaction of SIRK1 and QSK1

Project description:In this project, we want to search sucrose-induced interaction partners for LRR-receptor kinases SIRK1 and QSK1.

2019-11-12 | PXD011265 | Pride

Transcription profiling of Arabdiopsis wild type and G'4-3 seeds grown with different nitrate concentrations to investigate effects on seed dormancy

Project description:Freshly harvested Arabidopsis seeds (ecotype Col0 obtained under 10 mM nitrate nutrition, referred to as C10 seeds) are dormant and do not germinate when sown on an agarose-based medium. However, when C10 seeds are sown on 10mM nitrate or when Col0 mother plants are grown on 50 mM nitrate (referred to as C50 seeds), the produced seeds are non dormant and germinate readily on agarose. The G4-3 arabidopsis mutant which is impaired in nitrate assimilation and accumulates nitrate produces likewise non dormant seeds when grown on 10mM nitrate (referred to as G4-3 seeds). The goals of these experiments are to better understand the effects of nitrate feeding of mother plants on the produced seed dormancy.

2007-01-31 | E-MEXP-447 | biostudies-arrayexpress

New Insights into the Biosynthesis and Regulation of Wheat Amylose and Amylopectin from Proteomic and Phosphoproteomic Characterization of Granule-binding Proteins

Project description:Waxy starch has an important influence on bread dough and the qualities of breads. Generally, grain weight and yield in waxy wheat (Triticum aestivum L.) are significantly lower than in bread wheat. In this study, we performed the first proteomic and phosphoproteomic analyses of starch granule-binding proteins by comparing the waxy wheat cultivar Shannong 119 and the bread wheat cultivar Nongda 5181. The waxy and non-waxy wheats had similar starch granule morphological features and developmental patterns, and similar amylopectin quality in the grain. These results indicate that reduced amylose content does not affect amylopectin synthesis, but it causes significant reduction of total starch biosynthesis, grain size, weight and yield. Two-dimensional differential in-gel electrophoresis identified 40 differentially expressed protein (DEP) spots in waxy and non-waxy wheats, which belonged mainly to starch synthase (SS) I, SS IIa and granule-bound SS I. Most DEPs involved in amylopectin synthesis showed a similar expression pattern during grain development, suggesting relatively independent amylose and amylopectin synthesis pathways. Phosphoproteome analysis of starch granule-binding proteins, using TiO2 microcolumns and LC-MS/MS, showed that the total number of phosphoproteins and their phosphorylation levels in ND5181 were significantly higher than in SN119, but proteins controlling amylopectin synthesis had similar phosphorylation levels. Dynamic transcriptional expression profiling of starch biosynthesis-related genes indicated similar transcriptional expression profiles in both cultivars. Our results revealed that phosphorylation modifications played critical roles in amylose and amylopectin biosynthesis, but the lack of amylose did not affect the expression and phosphorylation of the starch granule-binding proteins involved in amylopectin biosynthesis.

2017-10-30 | PXD002805 | Pride

Human protein phosphorylation analysis

Project description:Mycobacterium tuberculosis (Mtb) protein kinase G (PknG) is a critical protein which can inhibit the maturation of phagosome and its fusion with the lysosome, providing Mtb’s latency and survival in the host. However, its signaling pathways and functions in the human body are still unclear and needed to be further studied. In our research, we used the human protein microarray to study the proteins interacting with PknG and have found a list of proteins that are potentially related to PknG in the host, which may contribute to the infection of Mtb. Among these positive hits, we found human protein CypA has strong interaction with PknG. We further validated the interaction between PknG and CypA in vitro and in vivo and investigated its role in the infection of macrophage cells. Finally, we found that PknG can significantly down-regulate protein expression level of CypA through phosphorylation in site of T107 as well as its induction of inflammatory response through NF-κB and ERK1/2 pathways. Meanwhile, the survival of Mycobacterium smegmetis (Msm) in the macrophages with overexpressing PknG is also improved with lessen of cytokines expression. Our results provided a resourceful research base of PknG in regards to various target human proteins and offered an insightful understanding of PknG’s function in the host.

2018-10-10 | PXD009237 | Pride

Perception of the peptide hormone Systemin

Project description:Cell suspension cultures of Solanum peruvianum were subjected to stimulation with 10nM systemin, the inactive systemin analog10nM a17, or injected with equal volume of water. For each treatment, a batch of cells was harvested after 2 minutes, 5 minutes, 15 minutes and 45 minutes. Untreated cells were harvested as controls. All treatments and time points were sampled as three independent replicates using independent sets of cells. Microsomal membranes were isolated from each sample, digested with trypsin, enriched for phosphopeptides and subjected to untargeted data-dependent acquisition by LC-MS/MS. Data analysis was performed jointly for all samples, and intensity values of triplicates were averaged.

2019-06-17 | PXD010819 | Pride

arcole / pgpr-Nitrogenous nutrition efficiency of canola in spring : identification of targets for plant breeding

Project description:pc_arcole - arcole / pgpr - What are the genes implicated in the efficiency of nitrogenous nutrition when A.thaliana is inoculated with a PGPR (Plant Growth Promoting Rhizobacteria)? - A.thaliana seeds germinated and grew during ten days until they were transfered in 6 different media: 0,5 mM nitrate with PGPR (Plant Growth Promoting Rhizobacteria), 0,5mM nitrate without PGPR, 2mM nitrate with PGPR, 2mM nitrate without PGPR, 20 mM nitrate with PGPR, 20 mM nitrate without PGPR. Young plantlets grew 7 days in these new mediums. Shoots are collected in eppendorf. 6 dye-swap - dose response,organ comparison,treated vs untreated comparison

2012-05-31 | E-GEOD-36730 | biostudies-arrayexpress