RRC ID 37014
著者 Nishimura K, Ishikawa S, Matsunami E, Yamauchi J, Homma K, Faulkner C, Oparka K, Jisaka M, Nagaya T, Yokota K, Nakagawa T.
タイトル New Gateway-compatible vectors for a high-throughput protein-protein interaction analysis by a bimolecular fluorescence complementation (BiFC) assay in plants and their application to a plant clathrin structure analysis.
ジャーナル Biosci Biotechnol Biochem
Abstract Protein-protein interactions (PPI) play key roles in various biological processes. The bimolecular fluorescence complementation (BiFC) assay is an excellent tool for routine PPI analyses in living cells. We developed new Gateway vectors for a high-throughput BiFC analysis of plants, adopting a monomeric Venus split just after the tenth β-strand, and analyzed the interaction between Arabidopsis thaliana coated vesicle coatmers, the clathrin heavy chain (CHC), and the clathrin light chain (CLC). In competitive BiFC tests, CLC interacted with CHC through a coiled-coil motif in the middle section of CLC. R1340, R1448, and K1512 in CHC and W94 in CLC are potentially key amino acids underlying the inter-chain interaction, consistent with analyses based on homology modeling. Our Gateway BiFC system, the V10-BiFC system, provides a useful tool for a PPI analysis in living plant cells. The CLC-CHC interaction identified may facilitate clathrin triskelion assembly needed for cage formation.
巻・号 79(12)
ページ 1995-2006
公開日 2015-1-1
DOI 10.1080/09168451.2015.1060847
PMID 26193449
MeSH Amino Acid Sequence Arabidopsis / metabolism* Arabidopsis Proteins / chemistry Arabidopsis Proteins / genetics Arabidopsis Proteins / metabolism* Clathrin / chemistry Clathrin / genetics Clathrin / metabolism* Genetic Vectors / genetics* Models, Molecular Molecular Sequence Data Mutation Protein Conformation Protein Interaction Mapping / methods* Sequence Homology, Amino Acid Spectrometry, Fluorescence
IF 1.516
引用数 7
WOS 分野 FOOD SCIENCE & TECHNOLOGY BIOTECHNOLOGY & APPLIED MICROBIOLOGY CHEMISTRY, APPLIED BIOCHEMISTRY & MOLECULAR BIOLOGY
リソース情報
シロイヌナズナ / 植物培養細胞・遺伝子 pda20053