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

Authors: Zábrady, Matej; Hrdinová, Vendula; Müller, Bruno; Conrad, Udo; Hejátko, Jan; Janda, Lubomír;

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.

Keywords

Science, Arabidopsis, 1100 General Agricultural and Biological Sciences, 580 Plants (Botany), Antibodies, Cytosol, 10126 Department of Plant and Microbial Biology, 1300 General Biochemistry, Genetics and Molecular Biology, Gene Expression Regulation, Plant, Gene Silencing, Protein Interaction Maps, 10211 Zurich-Basel Plant Science Center, 1000 Multidisciplinary, Arabidopsis Proteins, Q, Phosphotransferases, R, Recombinant Proteins, Medicine, Protein Kinases, Research Article, Signal Transduction

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    popularity
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    influence
    This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
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    impulse
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selected citations
These citations are derived from selected sources.
This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Citations provided by BIP!
popularity
This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
BIP!Popularity provided by BIP!
influence
This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Influence provided by BIP!
impulse
This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
BIP!Impulse provided by BIP!
8
Average
Average
Average
Green
gold