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Poster-17

Fluorescent probes of Aspergillus oryzae lectin fused with fluorescent proteins are internalized into golgi via endocytosis

초록

영어

The a(1,6)-fucose attached to the core N-glycan (core fucose) of glycoproteins has been known to play essential roles in various pathophysiological events, including oncogenesis and metastasis. Aspergillus oryzae lectin (AOL) encoded by the fleA gene has been reported to bind to N-glycans containing this core fucose. In our previous study, the fleA gene was fused with genes of fluorescent proteins to generate fluorescent probes. Their binding capabilities to core fucose was confirmed by lectin blot and cytochemical staining. Here, we describe the internalization properties of the FleA-fluorescent fusion proteins. During cell imaging experiments, we surprisingly found that these proteins could be internalized into intracellular compartments, especially Golgi apparatus, when applied to live cells. In contrast, their fluorescences were observed only on the cell surface when applied to the fixed cells or the metabolically arrested cells at low temperature. These results suggested that FleA-fluorescent fusion proteins first bound to the core-fucosylated glycoprotein located on the cell surface and then could be internalized by the active cellular process. This internalization was shown to occur depending on a clathrin-mediated pathway by endocytosis inhibitor assay. Taken together, FleA-fluorescent fusion proteins can be employed as a vesicle for delivery of useful materials to the inside of cells as well as a valuable fluorescent probe for detection of core fucose.

저자정보

  • Ji-Young Mun Integrative Omics Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Biosystems and Bioengineering Program, School of Science, University of Science and Technology (UST)
  • Yu Jin Kim Integrative Omics Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)
  • Kyung Jin Lee Integrative Omics Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)
  • Min Sun Sung Integrative Omics Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)
  • Ohsuk Kwon Integrative Omics Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Biosystems and Bioengineering Program, School of Science, University of Science and Technology (UST)
  • Doo-Byoung Oh Integrative Omics Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Biosystems and Bioengineering Program, School of Science, University of Science and Technology (UST)

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