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Identification of flagellin peptides in N. benthamiana apoplastic fluid

ABSTRACT: Purified flagellin from Pseudomonas Pta6605 WT was supplemented with N. benthamiana apoplastic fluids from plants transiently expressing only p19 or p19 and bGAL1 (a galactosidase). By performing subsequent LC/MSMS we wanted to identify and quantify the levels of flagellin peptides released by endogenous peptidases in the AF.

INSTRUMENT(S): LTQ Orbitrap Elite

ORGANISM(S): Nicotiana Benthamiana Pseudomonas Amygdali Pv. Tabaci

SUBMITTER: Farnusch Kaschani

LAB HEAD: Farnusch Kaschani

PROVIDER: PXD011823 | Pride | 2019-04-15

REPOSITORIES: Pride

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Glycosidase and glycan polymorphism control hydrolytic release of immunogenic flagellin peptides.

Buscaill Pierre P Chandrasekar Balakumaran B Sanguankiattichai Nattapong N Kourelis Jiorgos J Kaschani Farnusch F Thomas Emma L EL Morimoto Kyoko K Kaiser Markus M Preston Gail M GM Ichinose Yuki Y van der Hoorn Renier A L RAL

Science (New York, N.Y.) 20190401 6436

Plants and animals recognize conserved flagellin fragments as a signature of bacterial invasion. These immunogenic elicitor peptides are embedded in the flagellin polymer and require hydrolytic release before they can activate cell surface receptors. Although much of flagellin signaling is understood, little is known about the release of immunogenic fragments. We discovered that plant-secreted β-galactosidase 1 (BGAL1) of <i>Nicotiana benthamiana</i> promotes hydrolytic elicitor release and acts ...[more]

PMID: 30975858

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Re-annotated role of beta-Galactosidase in Flagellin deglycosylation

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2019-04-15 | PXD010461 | Pride

The transcriptome, extracellular proteome and active secretome of agroinfiltrated N. benthamiana uncover an expanded protease repertoire

Project description:Background: The infiltration of disarmed Agrobacterium tumefaciens into leaves of Nicotiana benthamiana (agroinfiltration) is a widely-adopted procedure to produce and study proteins in plant science, for instance to determine subcellular localization and protein-protein interactions. Agroinfiltration also facilitates quick and safe production of biopharmaceuticals. However, the agroinfiltration expression platform is hampered by unintended proteolysis, some of which might be associated with an elevated immune response. Results: Here, we studied the transcriptome, extracellular proteome and active secretome of agroinfiltrated leaves over a time course, with and without the p19 silencing inhibitor. Remarkably, p19 expression had little effect on the leaf transcriptome and no effect on the extracellular proteome. 25% of the detected transcripts changed in abundance upon agroinfiltration, associated with a gradual upregulation of immunity and a downregulation of photosynthesis. By contrast, 70% of the extracellular proteins increase in abundance and for some proteins this coincides with increased efficiency of extracellular delivery. The protease repertoire of N. benthamiana is expanded compared to other model plants and consists of 1245 proteases and protease homologs. We detect transcripts encoding for 975 proteases, identified extracellular peptides from 196 proteases and monitored 17 active extracellular Ser and Cys proteases in agroinfiltrated leaves. Conclusions: Analysis of agroinfiltrated leaves at transcript, protein and activity levels uncover transcriptional and post-transcriptional/translational reprogramming upon agroinfiltration, associated with substantial increase of the extracellular proteome, and a complex, expanded protease repertoire. This dataset increases our understanding of the plant response to agroinfiltration and indicates ways to improve a key expression platform for both plant science and molecular farming.

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Ubiquitin-independent disassembly by a p97 AAA-ATPase complex drives PP1 holoenzyme formation

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2019-05-06 | PXD011588 | Pride

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2017-02-22 | PXD005266 | Pride

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2018-05-17 | PXD008538 | Pride

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