title: Studying interactions between chloroplast proteins in intact plant cells using bimolecular fluorescence complementation and Förster resonance energy transfer.
abstract: Protein-protein interactions play crucial roles in the execution of many cellular functions, including those in plastids. Identifying and characterising protein-protein interactions can yield valuable information regarding the function of a protein and can also contribute towards understanding protein-protein interaction networks in plastids, thereby contributing to a better understanding of cellular processes. Here, we describe the planning and experimental procedures required to perform both bimolecular fluorescence complementation and Förster resonance energy transfer assays to detect protein-protein interactions. Arabidopsis is well-suited for microscopy and its small size facilitates live cell imaging, enabling observation of protein-protein interactions in living chloroplasts. The methods described in this chapter can be used to analyse protein-protein interactions of two known proteins and to dissect interacting protein domains.
authors.affiliation: Centre For Organelle Research, Universitetet i Stavanger, Stavanger, Norway.
authors.forename: Jodi
authors.initials: J
authors.lastname: Maple
authors.forename: Simon G
authors.initials: SG
authors.lastname: Møller
mesh_terms: D016960:Agrobacterium tumefaciens / Q000235:genetics; D017360:Arabidopsis / Q000166:cytology* / Q000378:metabolism*; D060365:Chloroplast Proteins / Q000235:genetics / Q000378:metabolism*; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics; D008853:Microscopy; D018515:Plant Leaves / Q000166:cytology / Q000378:metabolism; D025941:Protein Interaction Mapping / Q000379:methods*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D014026:Nicotiana / Q000166:cytology* / Q000378:metabolism*; D014170:Transformation, Genetic
chemical_list: D060365:Chloroplast Proteins; D011993:Recombinant Fusion Proteins