title: Isolation of protein complexes from the model legume Medicago truncatula by tandem affinity purification in hairy root cultures.
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.
authors.affiliation: Department of Plant Systems Biology, VIB, Technologiepark 927, B-9052, Gent, Belgium.|Department of Plant Biotechnology and Bioinformatics, Ghent University, Technologiepark 927, B-9052, Gent, Belgium.
authors.forename: Jonas
authors.initials: J
authors.lastname: Goossens
authors.affiliation: Department of Plant Systems Biology, VIB, Technologiepark 927, B-9052, Gent, Belgium.|Department of Plant Biotechnology and Bioinformatics, Ghent University, Technologiepark 927, B-9052, Gent, Belgium.
authors.forename: Nathan
authors.initials: N
authors.lastname: De Geyter
authors.affiliation: Department of Plant Systems Biology, VIB, Technologiepark 927, B-9052, Gent, Belgium.|Department of Plant Biotechnology and Bioinformatics, Ghent University, Technologiepark 927, B-9052, Gent, Belgium.|Department of Medical Protein Research, VIB, Albert Baertsoenkaai 3, B-9000, Gent, Belgium.|Department of Biochemistry, Ghent University, Albert Baertsoenkaai 3, B-9000, Gent, Belgium.
authors.forename: Alan
authors.initials: A
authors.lastname: Walton
authors.affiliation: Department of Plant Systems Biology, VIB, Technologiepark 927, B-9052, Gent, Belgium.|Department of Plant Biotechnology and Bioinformatics, Ghent University, Technologiepark 927, B-9052, Gent, Belgium.
authors.forename: Dominique
authors.initials: D
authors.lastname: Eeckhout
authors.affiliation: Department of Plant Systems Biology, VIB, Technologiepark 927, B-9052, Gent, Belgium.|Department of Plant Biotechnology and Bioinformatics, Ghent University, Technologiepark 927, B-9052, Gent, Belgium.
authors.forename: Jan
authors.initials: J
authors.lastname: Mertens
authors.affiliation: Department of Plant Systems Biology, VIB, Technologiepark 927, B-9052, Gent, Belgium.|Department of Plant Biotechnology and Bioinformatics, Ghent University, Technologiepark 927, B-9052, Gent, Belgium.
authors.forename: Jacob
authors.initials: J
authors.lastname: Pollier
authors.affiliation: Department of Plant Systems Biology, VIB, Technologiepark 927, B-9052, Gent, Belgium.|Department of Plant Biotechnology and Bioinformatics, Ghent University, Technologiepark 927, B-9052, Gent, Belgium.
authors.forename: Jennifer
authors.initials: J
authors.lastname: Fiallos-Jurado
authors.affiliation: Department of Plant Systems Biology, VIB, Technologiepark 927, B-9052, Gent, Belgium.|Department of Plant Biotechnology and Bioinformatics, Ghent University, Technologiepark 927, B-9052, Gent, Belgium.
authors.forename: Annick
authors.initials: A
authors.lastname: De Keyser
authors.affiliation: Department of Plant Systems Biology, VIB, Technologiepark 927, B-9052, Gent, Belgium.|Department of Plant Biotechnology and Bioinformatics, Ghent University, Technologiepark 927, B-9052, Gent, Belgium.
authors.forename: Rebecca
authors.initials: R
authors.lastname: De Clercq
authors.affiliation: Department of Plant Systems Biology, VIB, Technologiepark 927, B-9052, Gent, Belgium.|Department of Plant Biotechnology and Bioinformatics, Ghent University, Technologiepark 927, B-9052, Gent, Belgium.
authors.forename: Jelle
authors.initials: J
authors.lastname: Van Leene
authors.affiliation: Department of Medical Protein Research, VIB, Albert Baertsoenkaai 3, B-9000, Gent, Belgium.|Department of Biochemistry, Ghent University, Albert Baertsoenkaai 3, B-9000, Gent, Belgium.
authors.forename: Kris
authors.initials: K
authors.lastname: Gevaert
authors.affiliation: Department of Plant Systems Biology, VIB, Technologiepark 927, B-9052, Gent, Belgium.|Department of Plant Biotechnology and Bioinformatics, Ghent University, Technologiepark 927, B-9052, Gent, Belgium.
authors.forename: Geert
authors.initials: G
authors.lastname: De Jaeger
authors.affiliation: Department of Plant Systems Biology, VIB, Technologiepark 927, B-9052, Gent, Belgium.|Department of Plant Biotechnology and Bioinformatics, Ghent University, Technologiepark 927, B-9052, Gent, Belgium.
authors.forename: Sofie
authors.initials: S
authors.lastname: Goormachtig
authors.affiliation: Department of Plant Systems Biology, VIB, Technologiepark 927, B-9052, Gent, Belgium.|Department of Plant Biotechnology and Bioinformatics, Ghent University, Technologiepark 927, B-9052, Gent, Belgium.
authors.forename: Alain
authors.initials: A
authors.lastname: Goossens
mesh_terms: D060054:Agrobacterium / Q000235:genetics; D046913:Medicago truncatula / Q000235:genetics / Q000378:metabolism*; D018517:Plant Roots / Q000235:genetics / Q000378:metabolism; D030821:Plants, Genetically Modified / Q000235:genetics / Q000378:metabolism*; D013559:Symbiosis / Q000235:genetics / Q000502:physiology
keywords: 
  Medicago truncatula
  ; CAF1; CKS1; JAZ1; hairy roots; legume; tandem affinity purification; technical advance