title: Development of fluorescent probes for the detection of fucosylated N-glycans using an Aspergillus oryzae lectin.
abstract: The α(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 core fucose. The fleA gene encoding AOL was cloned into an Escherichia coli expression vector and then fused with genes of fluorescent proteins for production of fusion proteins. The resulting FleA-fluorescent fusion proteins were expressed well in E. coli and shown to detect glycoproteins containing N-glycans with core fucose by lectin blot assay. It was also shown to bind to the surface of cancer cells highly expressing the fucosyltransferase VIII for attachment of core fucose. Surprisingly, we found that FleA-fluorescent fusion proteins could be internalized into the intracellular compartment, early endosome, when applied to live cells. This internalization was shown to occur through a clathrin-mediated pathway by endocytosis inhibitor assay. Taken together, these results suggest that FleA-fluorescent fusion proteins can be employed as a valuable fluorescent probe for the detection of fucosylated glycans and/or a useful vehicle for delivery of substances to the inside of cells.
authors: Integrative Omics Research Center, Korea Research Institute of Bioscience and Biotechnology, Yuseong-gu, Daejeon, South Korea.
authors: Ji-Young
authors: JY
authors: Mun
authors: Kyung Jin
authors: KJ
authors: Lee
authors: Yu Jin
authors: YJ
authors: Kim
authors: Ohsuk
authors: O
authors: Kwon
authors: Su-Jin
authors: SJ
authors: Kim
authors: Seung-Goo
authors: SG
authors: Lee
authors: Wei Sun
authors: WS
authors: Park
authors: Won Do
authors: WD
authors: Heo
authors: Doo-Byoung
authors: DB
authors: Oh
mesh_terms: D001236:Aspergillus oryzae / Q000235:genetics*; D004705:Endocytosis; D004926:Escherichia coli / Q000235:genetics; D005456:Fluorescent Dyes / Q000302:isolation & purification / Q000378:metabolism*; D005656:Fungal Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D037102:Lectins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D008164:Luminescent Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D011134:Polysaccharides / Q000032:analysis*; D011485:Protein Binding; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D005456:Fluorescent Dyes; D005656:Fungal Proteins; D037102:Lectins; D008164:Luminescent Proteins; D011134:Polysaccharides; D011993:Recombinant Fusion Proteins