title: Targeted in vivo inhibition of specific protein-protein interactions using recombinant antibodies.
abstract: With the growing availability of genomic sequence information, there is an increasing need for gene function analysis. Antibody-mediated "silencing" represents an intriguing alternative for the precise inhibition of a particular function of biomolecules. Here, we describe a method for selecting recombinant antibodies with a specific purpose in mind, which is to inhibit intrinsic protein-protein interactions in the cytosol of plant cells. Experimental procedures were designed for conveniently evaluating desired properties of recombinant antibodies in consecutive steps. Our selection method was successfully used to develop a recombinant antibody inhibiting the interaction of ARABIDOPSIS HISTIDINE PHOSPHOTRANSFER PROTEIN 3 with such of its upstream interaction partners as the receiver domain of CYTOKININ INDEPENDENT HISTIDINE KINASE 1. The specific down-regulation of the cytokinin signaling pathway in vivo demonstrates the validity of our approach. This selection method can serve as a prototype for developing unique recombinant antibodies able to interfere with virtually any biomolecule in the living cell.
authors.affiliation: Central European Institute of Technology, Masaryk University, Brno, Czech Republic.
authors.forename: Matej
authors.initials: M
authors.lastname: Zábrady
authors.affiliation: Central European Institute of Technology, Masaryk University, Brno, Czech Republic.
authors.forename: Vendula
authors.initials: V
authors.lastname: Hrdinová
authors.affiliation: Institute of Plant Biology & Zürich-Basel Plant Science Center, University of Zürich, Zürich, Switzerland.
authors.forename: Bruno
authors.initials: B
authors.lastname: Müller
authors.affiliation: Department of Molecular Genetics, Leibniz Institute of Plant Genetics and Crop Plant Research (IPK), Gatersleben, Germany.
authors.forename: Udo
authors.initials: U
authors.lastname: Conrad
authors.affiliation: Central European Institute of Technology, Masaryk University, Brno, Czech Republic.
authors.forename: Jan
authors.initials: J
authors.lastname: Hejátko
authors.affiliation: Central European Institute of Technology, Masaryk University, Brno, Czech Republic.
authors.forename: Lubomír
authors.initials: L
authors.lastname: Janda
mesh_terms: D000906:Antibodies / Q000008:administration & dosage / Q000276:immunology; D017360:Arabidopsis / Q000235:genetics; D029681:Arabidopsis Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology; D003600:Cytosol / Q000276:immunology / Q000378:metabolism*; D018506:Gene Expression Regulation, Plant; D020868:Gene Silencing / Q000276:immunology; D010770:Phosphotransferases / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology; D060066:Protein Interaction Maps / Q000235:genetics* / Q000276:immunology; D011494:Protein Kinases / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology; D011994:Recombinant Proteins / Q000008:administration & dosage / Q000276:immunology; D015398:Signal Transduction
chemical_list: C117968:AHP3 protein, Arabidopsis; D000906:Antibodies; D029681:Arabidopsis Proteins; D011994:Recombinant Proteins; D010770:Phosphotransferases; D011494:Protein Kinases; C101990:CKI1 protein, Arabidopsis