Development of fluorescent probes for the detection of fucosylated N-glycans using an Aspergillus oryzae lectin

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Title
Development of fluorescent probes for the detection of fucosylated N-glycans using an Aspergillus oryzae lectin
Author(s)
Ji Young Mun; Kyung Jin Lee; Y J Kim; Oh Suk Kwon; Su Jin Kim; Seung Goo Lee; W S Park; W D Heo; Doo-Byoung Oh
Bibliographic Citation
Applied Microbiology and Biotechnology, vol. 93, no. 1, pp. 251-260
Publication Year
2012
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.
Keyword
AOLClathrin-mediatedCore fucoseEndocytosisFluorescent proteins
ISSN
0175-7598
Publisher
Springer
DOI
http://dx.doi.org/10.1007/s00253-011-3549-z
Type
Article
Appears in Collections:
Division of Bio Technology Innovation > SME Support Center > 1. Journal Articles
Division of Biomaterials Research > 1. Journal Articles
Division of Research on National Challenges > Environmental diseases research center > 1. Journal Articles
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