Development of Fluorogenic Substrates of α- L -Fucosidase Useful for Inhibitor Screening and Gene-expression Profiling

Kazuki Miura, Takumi Tsukagoshi, Takako Hirano, Toshiyuki Nishio, Wataru Hakamata

Research output: Contribution to journalArticle

1 Citation (Scopus)


Inhibitors of human α-l-fucosidases, tissue α-l-fucosidase (tFuc), and plasma α-l-fucosidase reportedly play roles in multiple diseases, suggesting their therapeutic potential for gastric disease associated with Helicobacter pylori and fucosidosis. Terminal fucose linkages on glycoproteins and glycolipids are a natural substrate for both enzymes; however, there are currently no fluorogenic substrates allowing their cellular evaluation. Here, we described the development of novel three-color fluorogenic substrates for lysosome-localized tFuc that exhibited excellent specificity and sensitivity in three human cell lines. Additionally, we developed a cell-based high-throughput inhibitor screening system in a 96-well format and a cell-based inhibitory activity evaluation system in a 6-well format for tFuc inhibitors using this substrate, which allowed accurate quantification of the inhibition rate. Moreover, analysis of significant changes in gene expression resulting from 30% inhibition of tFuc in HeLa cells revealed potential roles in gastric disease.

Original languageEnglish
JournalACS Medicinal Chemistry Letters
Publication statusAccepted/In press - 2019 Jan 1
Externally publishedYes



  • DNA microarray
  • Fluorogenic substrate
  • High-throughput screening
  • Inhibitor
  • α- l -Fucosidase

ASJC Scopus subject areas

  • Biochemistry
  • Drug Discovery
  • Organic Chemistry

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