한빛사 논문
Abstract
Kyungtae Kanga,b,c,1, Sunghoon Jood,1, Ji Yu Choia,b,c,1, Sujeong Geumb,c, Seok-Pyo Honga,b,c, Seung-Yeul Leee,f, Yong Ho Kimf, Seong-Min Kimg, Myung-Han Yoong, Yoonkey Namd,2, Kyung-Bok Leee,2, Hee-Yoon Leeb,c,2, and Insung S. Choia,b,c,d,2
aCenter for Cell-Encapsulation Research,
bMolecular-Level Interface Research Center, and
Departments of cChemistry and
dBio and Brain Engineering, Korea Advanced Institute of Science and Technology (KAIST), Daejeon 305-701, Korea;
eDivision of Life Science, Korea Basic Science Institute, Daejeon 305-806, Korea;
fSKKU Advanced Institute of Nanotechnology, Sungkyunkwan University, Suwon 440-746, Korea; and
gSchool of Materials Science and Engineering, Gwangju Institute of Science and Technology, Gwangju 500-712, Korea
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.
metabolic labeling, bioorthogonal reaction, polysialic acid, 1,3-dipolar cycloaddition, primary hippocampal neuron
1K.K., S.J., and J.Y.C. contributed equally to this work.
2To whom correspondence may be addressed.
Author contributions: K.K., S.J., and I.S.C. designed research; K.K., S.J., and J.Y.C. performed research; S.G., S.-P.H., S.-Y.L., Y.H.K., and S.-M.K. contributed new reagents/analytic tools; K.K., S.J., J.Y.C., M.-H.Y., Y.N., K.-B.L., H.-Y.L., and I.S.C. analyzed data; and K.K., S.J., J.Y.C., and I.S.C. wrote the paper.
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