title: Endoplasmic Reticulum (ER)-Targeted, Galectin-Mediated Retrograde Transport by Using a HaloTag Carrier Protein.
abstract: Investigations into metabolic processes within the cell have often relied on genetic methods such as forced expression and knockout or knockdown techniques. An alternative approach would be introducing a molecule into the desired location inside the cell. To translocate compounds from outside cells into the endoplasmic reticulum (ER), we constructed a delivery carrier protein. This comprised N-terminal galectin-1 for cell-surface binding (G1), a protease cleavable sequence (ps), a HaloTag domain for attaching exogenous compounds (Halo), and a C-terminal KDEL sequence for ER retention. Fluorescently labeled G1-ps-Halo-KDEL passed through the Golgi apparatus and reached the ER. By using Man9 GlcNAc2 -BODIPY as a cargo compound, the carrier protein was also delivered into the ER with concomitant processing of mannose to Man5,6, by the ER-resident α1,2-mannosidase. G1-ps-Halo-KDEL might serve as a new type of delivery carrier protein to direct compounds into the ER.
authors.affiliation: ERATO, Science and Technology Agency (JST), 2-1 Hirosawa, Wako, Saitama, 351-0198, Japan.|College of Pharmacy, Korea University, 2511 Sejong-ro, Jochiwon-eup, Sejong, 339-700, South Korea.
authors.forename: Sang-Hyun
authors.initials: SH
authors.lastname: Son
authors.affiliation: ERATO, Science and Technology Agency (JST), 2-1 Hirosawa, Wako, Saitama, 351-0198, Japan.
authors.forename: Akira
authors.initials: A
authors.lastname: Seko
authors.affiliation: ERATO, Science and Technology Agency (JST), 2-1 Hirosawa, Wako, Saitama, 351-0198, Japan.
authors.forename: Shusaku
authors.initials: S
authors.lastname: Daikoku
authors.affiliation: ERATO, Science and Technology Agency (JST), 2-1 Hirosawa, Wako, Saitama, 351-0198, Japan.|Suntory Bioorganic Research Institute, 8-1-1 Seikadai, Seika-cho, Soraku-gun, Kyoto, 619-0284, Japan.
authors.forename: Kohki
authors.initials: K
authors.lastname: Fujikawa
authors.affiliation: ERATO, Science and Technology Agency (JST), 2-1 Hirosawa, Wako, Saitama, 351-0198, Japan.|Faculty of Pharmaceutical Sciences, Aomori University, 2-3-1 Kohbata, Aomori, 030-0943, Japan.
authors.forename: Katsuhiko
authors.initials: K
authors.lastname: Suzuki
authors.affiliation: ERATO, Science and Technology Agency (JST), 2-1 Hirosawa, Wako, Saitama, 351-0198, Japan.|Synthetic Cellular Chemistry Laboratory, RIKEN, 2-1 Hirosawa, Wako, Saitama, 351-0198, Japan.
authors.forename: Yukishige
authors.initials: Y
authors.lastname: Ito
authors.affiliation: Institute of Glycoscience, Tokai University, 4-1-1 Kitakaname, Hiratsuka, Kanagawa, 259-1292, Japan. kanie@tokai-u.jp.|ERATO, Science and Technology Agency (JST), 2-1 Hirosawa, Wako, Saitama, 351-0198, Japan. kanie@tokai-u.jp.
authors.forename: Osamu
authors.identifier: http://orcid.org/0000-0003-4147-6868
authors.initials: O
authors.lastname: Kanie
mesh_terms: D001692:Biological Transport; D001896:Boron Compounds / Q000737:chemistry; D002352:Carrier Proteins / Q000378:metabolism*; D016503:Drug Delivery Systems; D004721:Endoplasmic Reticulum / Q000737:chemistry*; D004926:Escherichia coli / Q000737:chemistry / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D037161:Galectins / Q000378:metabolism*; D006056:Golgi Apparatus / Q000737:chemistry; D008301:Maleimides / Q000737:chemistry / Q000378:metabolism; D011134:Polysaccharides / Q000737:chemistry / Q000378:metabolism
chemical_list: C095489:4,4-difluoro-4-bora-3a,4a-diaza-s-indacene; C504424:Alexa Fluor 488 C5-maleimide; D001896:Boron Compounds; D002352:Carrier Proteins; D037161:Galectins; D008301:Maleimides; D011134:Polysaccharides
keywords: HaloTag; drug delivery; endoplasmic reticulum; galectin; glycoproteins; retrograde delivery