Endoplasmic Reticulum (ER)‐Targeted,Galectin‐Mediated Retrograde Transport by Using a HaloTag Carrier Protein |
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| Authors: | Dr Sang‐Hyun Son Dr Akira Seko Dr Shusaku Daikoku Dr Kohki Fujikawa Assoc?Prof?Dr Katsuhiko Suzuki Chief?Scientist?Dr Yukishige Ito Prof?Dr Osamu Kanie |
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| Affiliation: | 1. ERATO, Science and Technology Agency (JST), Wako, Saitama, Japan;2. College of Pharmacy, Korea University, Jochiwon-eup, Sejong, South Korea;3. Suntory Bioorganic Research Institute, Seika-cho, Soraku-gun, Kyoto, Japan;4. Faculty of Pharmaceutical Sciences, Aomori University, Aomori, Japan;5. Synthetic Cellular Chemistry Laboratory, RIKEN, Wako, Saitama, Japan;6. Institute of Glycoscience, Tokai University, Hiratsuka, Kanagawa, Japan |
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| 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 Man9GlcNAc2‐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. |
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| Keywords: | drug delivery endoplasmic reticulum galectin glycoproteins HaloTag retrograde delivery |
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