Journal Article

Detection of <i>in vivo</i> protein interactions between Snf1-related kinase subunits with intron-tagged epitope-labelling in plants cells

Alejandro Ferrando, Zsuzsana Koncz-Kálmán, Rosa Farràs, Antonio Tiburcio, Jeff Schell and Csaba Koncz

in Nucleic Acids Research

Volume 29, issue 17, pages 3685-3693
Published in print September 2001 | ISSN: 0305-1048
Published online September 2001 | e-ISSN: 1362-4962 | DOI: http://dx.doi.org/10.1093/nar/29.17.3685
Detection of in vivo protein interactions between  Snf1-related kinase subunits with intron-tagged epitope-labelling in plants cells

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  • Chemistry
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  • Molecular and Cell Biology

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Plant orthologs of the yeast sucrose non-fermenting (Snf1) kinase and mammalian AMP-activated protein kinase (AMPK) represent an emerging class of important regulators of metabolic and stress signalling. The catalytic α-subunits of plant Snf1-related kinases (SnRKs) interact in the yeast two-hybrid system with different proteins that share conserved domains with the β- and γ-subunits of Snf1 and AMPKs. However, due to the lack of a robust technique allowing the detection of protein interactions in plant cells, it is unknown whether these proteins indeed occur in SnRK complexes in vivo. Here we describe a double-labelling technique, using intron-tagged hemagglutinin (HA) and c-Myc epitope sequences, which provides a simple tool for co-immunopurification of interacting proteins expressed in Agrobacterium-transformed Arabidopsis cells. This generally applicable plant protein interaction assay was used to demonstrate that AKINβ2, a plant ortholog of conserved Snf1/AMPK β-subunits, forms different complexes with the catalytic α-subunits of Arabidopsis SnRK protein kinases AKIN10 and AKIN11 in vivo.

Journal Article.  6036 words.  Illustrated.

Subjects: Chemistry ; Biochemistry ; Bioinformatics and Computational Biology ; Genetics and Genomics ; Molecular and Cell Biology

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