Tissue-based metabolic labeling of polysialic acids in living primary hippocampal neurons
| Autor: | Seung-Yeul Lee, Seong-Min Kim, Kyung-Bok Lee, Ji Yu Choi, Sunghoon Joo, Myung-Han Yoon, Yong Ho Kim, Yoonkey Nam, Insung S. Choi, Sujeong Geum, Hee-Yoon Lee, Kyungtae Kang, Seok-Pyo Hong |
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| Rok vydání: | 2015 |
| Předmět: |
Neurons
Proteomics Multidisciplinary Neurite Glycobiology Polysialic acid Nerve Tissue Proteins Biology Hippocampal formation Hippocampus Sialic acid Rats Rats Sprague-Dawley chemistry.chemical_compound medicine.anatomical_structure nervous system chemistry Biochemistry PNAS Plus medicine Sialic Acids Animals Neural cell adhesion molecule Neuron N-Acetylneuraminic acid |
| Zdroj: | Proceedings of the National Academy of Sciences of the United States of America. 112(3) |
| ISSN: | 1091-6490 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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