title: Tissue-based metabolic labeling of polysialic acids in living primary hippocampal neurons.
abstract: The posttranslational modification of neural cell-adhesion molecule (NCAM) with polysialic acid (PSA) and the spatiotemporal distribution of PSA-NCAM play an important role in the neuronal development. In this work, we developed a tissue-based strategy for metabolically incorporating an unnatural monosaccharide, peracetylated N-azidoacetyl-D-mannosamine, in the sialic acid biochemical pathway to present N-azidoacetyl sialic acid to PSA-NCAM. Although significant neurotoxicity was observed in the conventional metabolic labeling that used the dissociated neuron cells, neurotoxicity disappeared in this modified strategy, allowing for investigation of the temporal and spatial distributions of PSA in the primary hippocampal neurons. PSA-NCAM was synthesized and recycled continuously during neuronal development, and the two-color labeling showed that newly synthesized PSA-NCAMs were transported and inserted mainly to the growing neurites and not significantly to the cell body. This report suggests a reliable and cytocompatible method for in vitro analysis of glycans complementary to the conventional cell-based metabolic labeling for chemical glycobiology.
authors.affiliation: Center for Cell-Encapsulation Research, Molecular-Level Interface Research Center, and Departments of Chemistry and.
authors.forename: Kyungtae
authors.initials: K
authors.lastname: Kang
authors.affiliation: Bio and Brain Engineering, Korea Advanced Institute of Science and Technology (KAIST), Daejeon 305-701, Korea;
authors.forename: Sunghoon
authors.initials: S
authors.lastname: Joo
authors.affiliation: Center for Cell-Encapsulation Research, Molecular-Level Interface Research Center, and Departments of Chemistry and.
authors.forename: Ji Yu
authors.initials: JY
authors.lastname: Choi
authors.affiliation: Molecular-Level Interface Research Center, and Departments of Chemistry and.
authors.forename: Sujeong
authors.initials: S
authors.lastname: Geum
authors.affiliation: Center for Cell-Encapsulation Research, Molecular-Level Interface Research Center, and Departments of Chemistry and.
authors.forename: Seok-Pyo
authors.initials: SP
authors.lastname: Hong
authors.affiliation: Division of Life Science, Korea Basic Science Institute, Daejeon 305-806, Korea; SKKU Advanced Institute of Nanotechnology, Sungkyunkwan University, Suwon 440-746, Korea; and.
authors.forename: Seung-Yeul
authors.initials: SY
authors.lastname: Lee
authors.affiliation: SKKU Advanced Institute of Nanotechnology, Sungkyunkwan University, Suwon 440-746, Korea; and.
authors.forename: Yong Ho
authors.initials: YH
authors.lastname: Kim
authors.affiliation: School of Materials Science and Engineering, Gwangju Institute of Science and Technology, Gwangju 500-712, Korea.
authors.forename: Seong-Min
authors.initials: SM
authors.lastname: Kim
authors.affiliation: School of Materials Science and Engineering, Gwangju Institute of Science and Technology, Gwangju 500-712, Korea.
authors.forename: Myung-Han
authors.initials: MH
authors.lastname: Yoon
authors.affiliation: Bio and Brain Engineering, Korea Advanced Institute of Science and Technology (KAIST), Daejeon 305-701, Korea; ynam@kaist.ac.kr kblee@kbsi.re.kr leehy@kaist.ac.kr ischoi@kaist.ac.kr.
authors.forename: Yoonkey
authors.initials: Y
authors.lastname: Nam
authors.affiliation: Division of Life Science, Korea Basic Science Institute, Daejeon 305-806, Korea; ynam@kaist.ac.kr kblee@kbsi.re.kr leehy@kaist.ac.kr ischoi@kaist.ac.kr.
authors.forename: Kyung-Bok
authors.initials: KB
authors.lastname: Lee
authors.affiliation: Molecular-Level Interface Research Center, and Departments of Chemistry and ynam@kaist.ac.kr kblee@kbsi.re.kr leehy@kaist.ac.kr ischoi@kaist.ac.kr.
authors.forename: Hee-Yoon
authors.initials: HY
authors.lastname: Lee
authors.affiliation: Center for Cell-Encapsulation Research, Molecular-Level Interface Research Center, and Departments of Chemistry and Bio and Brain Engineering, Korea Advanced Institute of Science and Technology (KAIST), Daejeon 305-701, Korea; ynam@kaist.ac.kr kblee@kbsi.re.kr leehy@kaist.ac.kr ischoi@kaist.ac.kr.
authors.forename: Insung S
authors.initials: IS
authors.lastname: Choi
mesh_terms: D000818:Animals; D006624:Hippocampus / Q000166:cytology / Q000378:metabolism*; D009419:Nerve Tissue Proteins / Q000378:metabolism; D009474:Neurons / Q000378:metabolism*; D040901:Proteomics; D051381:Rats; D017207:Rats, Sprague-Dawley; D012794:Sialic Acids / Q000378:metabolism*
chemical_list: D009419:Nerve Tissue Proteins; D012794:Sialic Acids; C021319:polysialic acid
keywords: 1,3-dipolar cycloaddition; bioorthogonal reaction; metabolic labeling; polysialic acid; primary hippocampal neuron