title: Protein ligand-tethered synthetic calcium indicator for localization control and spatiotemporal calcium imaging in plant cells.
abstract: In plant biology, calcium ions are involved in a variety of intriguing biological phenomena as a secondary messenger. However, most conventional calcium indicators are not applicable for plant cells because of the difficulty with their localization control in plant cells. We here introduce a method to monitor spatiotemporal Ca(2+) dynamics in living plant cells based on linking the synthetic calcium indicator Calcium Green-1 to a natural product-based protein ligand. In a proof-of-concept study using cultured BY-2 cells overexpressing the target protein for the ligand, the ligand-tethered probe accumulated in the cytosol and nucleus, and enabled real-time monitoring of the cytosolic and nucleus Ca(2+) dynamics under the physiological condition. The present strategy using ligand-tethered fluorescent sensors may be successfully applied to reveal the spatiotemporal dynamics of calcium ions in living plant cells.
authors.affiliation: Department of Chemistry, Graduate School of Science, Tohoku University, 6-3 Aramaki-Aza-Aoba, Aoba-ku, Sendai 980-8578, Japan.
authors.forename: Yousuke
authors.initials: Y
authors.lastname: Takaoka
authors.affiliation: Department of Chemistry, Graduate School of Science, Tohoku University, 6-3 Aramaki-Aza-Aoba, Aoba-ku, Sendai 980-8578, Japan.
authors.forename: Miyuki
authors.initials: M
authors.lastname: Shigenaga
authors.affiliation: Department of Chemistry, Graduate School of Science, Tohoku University, 6-3 Aramaki-Aza-Aoba, Aoba-ku, Sendai 980-8578, Japan.
authors.forename: Masaki
authors.initials: M
authors.lastname: Imai
authors.affiliation: Department of Chemistry, Graduate School of Science, Tohoku University, 6-3 Aramaki-Aza-Aoba, Aoba-ku, Sendai 980-8578, Japan.
authors.forename: Yuuki
authors.initials: Y
authors.lastname: Nukadzuka
authors.affiliation: Department of Chemistry, Graduate School of Science, Tohoku University, 6-3 Aramaki-Aza-Aoba, Aoba-ku, Sendai 980-8578, Japan.
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Ishimaru
authors.affiliation: Laboratory of Plant Cell Wall Biology, Graduate School of Life Sciences, Tohoku University, 6-3 Aramaki-Aza-Aoba, Aoba-ku, Sendai 980-8578, Japan.
authors.forename: Kei
authors.initials: K
authors.lastname: Saito
authors.affiliation: Laboratory of Plant Cell Wall Biology, Graduate School of Life Sciences, Tohoku University, 6-3 Aramaki-Aza-Aoba, Aoba-ku, Sendai 980-8578, Japan.
authors.forename: Ryusuke
authors.initials: R
authors.lastname: Yokoyama
authors.affiliation: Laboratory of Plant Cell Wall Biology, Graduate School of Life Sciences, Tohoku University, 6-3 Aramaki-Aza-Aoba, Aoba-ku, Sendai 980-8578, Japan.
authors.forename: Kazuhiko
authors.initials: K
authors.lastname: Nishitani
authors.affiliation: Department of Chemistry, Graduate School of Science, Tohoku University, 6-3 Aramaki-Aza-Aoba, Aoba-ku, Sendai 980-8578, Japan. Electronic address: ueda@m.tohoku.ac.jp.
authors.forename: Minoru
authors.initials: M
authors.lastname: Ueda
mesh_terms: D002118:Calcium / Q000032:analysis* / Q000378:metabolism*; D002478:Cells, Cultured; D003600:Cytosol / Q000737:chemistry / Q000378:metabolism; D005456:Fluorescent Dyes / Q000032:analysis* / Q000737:chemistry; D008024:Ligands; D008856:Microscopy, Fluorescence; D015394:Molecular Structure; D009930:Organic Chemicals / Q000032:analysis / Q000138:chemical synthesis / Q000737:chemistry; D062211:Spatio-Temporal Analysis; D014026:Nicotiana / Q000166:cytology / Q000378:metabolism*
chemical_list: D005456:Fluorescent Dyes; D008024:Ligands; D009930:Organic Chemicals; C071336:calcium green; D002118:Calcium
keywords: Calcium indicator; Chemical biology; Natural products; Plant cell