title: Analysis of Ethylene Receptor Interactions by Co-immunoprecipitation Assays.
abstract: Ethylene receptors are predominantly localized to the endoplasmic reticulum (ER) membrane, and coordinate ethylene signal output through protein-protein interactions with each other and additional signaling components. Here, we describe a co-immunoprecipitation (Co-IP) assay based on the use of the Tandem Affinity Purification (TAP) tag to examine the interactions of ethylene receptors in plant extracts. Human IgG-agarose beads are used to pull down TAP-tagged versions of the protein of interest from detergent extracts of Arabidopsis membranes, and the precipitate then is analyzed immunologically for co-purification of the ethylene receptors. This method has been successfully used to examine interactions of the receptors with each other as well as with the Raf-like kinase CTR1.
authors.affiliation: State Key Laboratory of Hybrid Rice, College of Life Sciences, Wuhan University, 129 Luoyu Road, Wuhan, 430072, China. zygao@whu.edu.cn.
authors.forename: Zhiyong
authors.initials: Z
authors.lastname: Gao
authors.affiliation: Department of Biological Sciences, Dartmouth College, Hanover, NH, 03755, USA.
authors.forename: G Eric
authors.initials: GE
authors.lastname: Schaller
mesh_terms: D005030:Ethylenes / Q000378:metabolism*; D047468:Immunoprecipitation / Q000379:methods*; D010940:Plant Proteins / Q000378:metabolism*; D011485:Protein Binding; D011956:Receptors, Cell Surface / Q000378:metabolism*
chemical_list: D005030:Ethylenes; D010940:Plant Proteins; D011956:Receptors, Cell Surface; C083750:ethylene receptors, plant; C036216:ethylene
keywords: CTR1; Co-IP; Ethylene receptor; IgG-agarose; Protein a; Protein interaction; TAP-tag