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Isolation of protein complexes from the model legume Medicago truncatula by tandem affinity purification in hairy root cultures

Jonas Goossens (UGent) , Nathan De Geyter (UGent) , Alan Walton (UGent) , Dominique Eeckhout (UGent) , Jan Mertens (UGent) , Jacob Pollier (UGent) , Jennifer Liliana Fiallos Jurado, Annick De Keyser (UGent) , Rebecca De Clercq (UGent) , Jelle Van Leene (UGent) , et al.
(2016) PLANT JOURNAL. 88(3). p.476-489
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Abstract
Tandem affinity purification coupled to mass spectrometry (TAP-MS) is one of the most powerful techniques to isolate protein complexes and elucidate protein interaction networks. Here, we describe the development of a TAP-MS strategy for the model legume Medicago truncatula, which is widely studied for its ability to produce valuable natural products and to engage in endosymbiotic interactions. As biological material, transgenic hairy roots, generated through Agrobacterium rhizogenes-mediated transformation of M.truncatula seedlings, were used. As proof of concept, proteins involved in the cell cycle, transcript processing and jasmonate signalling were chosen as bait proteins, resulting in a list of putative interactors, many of which confirm the interologue concept of protein interactions, and which can contribute to biological information about the functioning of these bait proteins in planta. Subsequently, binary protein-protein interactions among baits and preys, and among preys were confirmed by a systematic yeast two-hybrid screen. Together, by establishing a M.truncatula TAP-MS platform, we extended the molecular toolbox of this model species.
Keywords
Medicago truncatula, tandem affinity purification, legume, hairy roots, JAZ1, CKS1, CAF1, technical advance, CCR4-NOT COMPLEX, MASS-SPECTROMETRY, ARABIDOPSIS-THALIANA, SACCHAROMYCES-CEREVISIAE, SYMBIOTIC ASSOCIATIONS, INTERACTION PROTEOMICS, ARBUSCULAR MYCORRHIZA, TRANSFORMED ROOTS, GENE-EXPRESSION, NUCLEASE MODULE

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Chicago
Goossens, Jonas, Nathan De Geyter, Alan Walton, Dominique Eeckhout, Jan Mertens, Jacob Pollier, Jennifer Liliana Fiallos Jurado, et al. 2016. “Isolation of Protein Complexes from the Model Legume Medicago Truncatula by Tandem Affinity Purification in Hairy Root Cultures.” Plant Journal 88 (3): 476–489.
APA
Goossens, Jonas, De Geyter, N., Walton, A., Eeckhout, D., Mertens, J., Pollier, J., Fiallos Jurado, J. L., et al. (2016). Isolation of protein complexes from the model legume Medicago truncatula by tandem affinity purification in hairy root cultures. PLANT JOURNAL, 88(3), 476–489.
Vancouver
1.
Goossens J, De Geyter N, Walton A, Eeckhout D, Mertens J, Pollier J, et al. Isolation of protein complexes from the model legume Medicago truncatula by tandem affinity purification in hairy root cultures. PLANT JOURNAL. 2016;88(3):476–89.
MLA
Goossens, Jonas, Nathan De Geyter, Alan Walton, et al. “Isolation of Protein Complexes from the Model Legume Medicago Truncatula by Tandem Affinity Purification in Hairy Root Cultures.” PLANT JOURNAL 88.3 (2016): 476–489. Print.
@article{8500649,
  abstract     = {Tandem affinity purification coupled to mass spectrometry (TAP-MS) is one of the most powerful techniques to isolate protein complexes and elucidate protein interaction networks. Here, we describe the development of a TAP-MS strategy for the model legume Medicago truncatula, which is widely studied for its ability to produce valuable natural products and to engage in endosymbiotic interactions. As biological material, transgenic hairy roots, generated through Agrobacterium rhizogenes-mediated transformation of M.truncatula seedlings, were used. As proof of concept, proteins involved in the cell cycle, transcript processing and jasmonate signalling were chosen as bait proteins, resulting in a list of putative interactors, many of which confirm the interologue concept of protein interactions, and which can contribute to biological information about the functioning of these bait proteins in planta. Subsequently, binary protein-protein interactions among baits and preys, and among preys were confirmed by a systematic yeast two-hybrid screen. Together, by establishing a M.truncatula TAP-MS platform, we extended the molecular toolbox of this model species.},
  author       = {Goossens, Jonas and De Geyter, Nathan and Walton, Alan and Eeckhout, Dominique and Mertens, Jan and Pollier, Jacob and Fiallos Jurado, Jennifer Liliana and De Keyser, Annick and De Clercq, Rebecca and Van Leene, Jelle and Gevaert, Kris and De Jaeger, Geert and Goormachtig, Sofie and Goossens, Alain},
  issn         = {0960-7412},
  journal      = {PLANT JOURNAL},
  keyword      = {Medicago truncatula,tandem affinity purification,legume,hairy roots,JAZ1,CKS1,CAF1,technical advance,CCR4-NOT COMPLEX,MASS-SPECTROMETRY,ARABIDOPSIS-THALIANA,SACCHAROMYCES-CEREVISIAE,SYMBIOTIC ASSOCIATIONS,INTERACTION PROTEOMICS,ARBUSCULAR MYCORRHIZA,TRANSFORMED ROOTS,GENE-EXPRESSION,NUCLEASE MODULE},
  language     = {eng},
  number       = {3},
  pages        = {476--489},
  title        = {Isolation of protein complexes from the model legume Medicago truncatula by tandem affinity purification in hairy root cultures},
  url          = {http://dx.doi.org/10.1111/tpj.13258},
  volume       = {88},
  year         = {2016},
}

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