Targeted In Vivo Inhibition of Specific Protein–Protein Interactions Using Recombinant Antibodies

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This publication doesn't include Faculty of Economics and Administration. It includes Central European Institute of Technology. Official publication website can be found on muni.cz.
Authors

ZÁBRADY Matej HRDINOVÁ Vendula MÜLLER Bruno CONRAD Udo HEJÁTKO Jan JANDA Lubomír

Year of publication 2014
Type Article in Periodical
Magazine / Source PLOS ONE : Public Library of Science
MU Faculty or unit

Central European Institute of Technology

Citation
Web http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0109875
Doi http://dx.doi.org/10.1371/journal.pone.0109875
Field Genetics and molecular biology
Keywords recombinant antibodies;immunomodulation;inhibition;protein-protein interactions;cytokinin signaling pathway
Description 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.
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