title: A novel pair of split venus fragments to detect protein-protein interactions by in vitro and in vivo bimolecular fluorescence complementation assays.
abstract: Protein-protein interactions are critical components of almost every cellular process. The bimolecular fluorescence complementation (BiFC) method has been used to detect protein-protein interactions in both living cells and cell-free systems. The BiFC method is based on the principle that a fluorescent protein is reassembled from its two complementary non-fluorescent fragments when an interaction occurs between two proteins, each one fused to each fragment. In vivo and in vitro BiFC assays, which use a new pair of split Venus fragments composed of VN210 (amino acids 1-210) and VC210 (amino acids 210-238), are useful tools to detect and quantify various protein-protein interactions (including the cofilin-actin and Ras-Raf interactions) with high specificity and low background fluorescence. Moreover, these assays can be applied to screen small-molecule inhibitors of protein-protein interactions.
authors.affiliation: Graduate School of Life Sciences, Tohoku University, Aramaki-aza-aoba, Aoba-ku, Sendai, Miyagi, 980-8578, Japan, kohashi@biology.tohoku.ac.jp.
authors.forename: Kazumasa
authors.initials: K
authors.lastname: Ohashi
authors.forename: Kensaku
authors.initials: K
authors.lastname: Mizuno
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D055808:Drug Discovery; D015870:Gene Expression; D057166:High-Throughput Screening Assays; D006801:Humans; D008164:Luminescent Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008856:Microscopy, Fluorescence / Q000379:methods; D010957:Plasmids / Q000235:genetics; D011485:Protein Binding / Q000187:drug effects; D025941:Protein Interaction Mapping / Q000379:methods; D011506:Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D008164:Luminescent Proteins; D011506:Proteins; D011994:Recombinant Proteins; C054562:yellow fluorescent protein, Bacteria