PMID- 32475856
OWN - NLM
STAT- MEDLINE
DCOM- 20210111
LR  - 20210111
IS  - 1347-5223 (Electronic)
IS  - 0009-2363 (Linking)
VI  - 68
IP  - 6
DP  - 2020
TI  - Development of Specific Fluorogenic Substrates for Human 
      beta-N-Acetyl-D-hexosaminidase A for Cell-Based Assays.
PG  - 526-533
LID - 10.1248/cpb.c20-00069 [doi]
AB  - Inhibitors of human beta-N-acetyl-D-hexosaminidase (hHEX) A and human O-GlcNAcase 
      (hOGA) reportedly play roles in multiple diseases, suggesting their potential for 
      pharmacological chaperone (PC) therapy of Sandhoff disease (SD) and Tay-Sachs 
      disease (TSD), as lysosomal storage diseases, and Alzheimer's disease and 
      progressive supranuclear palsy, respectively. In particular, hHEXA inhibitors as 
      PCs have been shown to successfully enhance hHEXA levels, leading to the chronic 
      form of SD and TSD. In the diagnosis of enzyme deficiencies in SD and TSD, 
      artificial hHEXA substrates based on 4-methylumbelliferone as a fluorophore are 
      available and generally used; however, they do not have sufficient performance to 
      screen for potential inhibitors for a PC therapy from compound libraries. 
      Further, there are currently few fluorogenic substrates for hHEXA suitable for 
      such requirements and there are no substrates ideal for cell-based inhibitor 
      screening. Here, we clarified the difference in enzyme active site structure 
      between hHEXA and hOGA from their tertiary structures. To develop 
      lysosome-localized hHEXA-specific fluorogenic substrates based on the difference 
      in their active site structures, our developed quinone methide cleavage substrate 
      design platform was applied for the molecular design of substrates. Thereafter, 
      we synthesized via the shortest route and evaluated novel three-color fluorogenic 
      substrates for hHEXA that exhibited excellent specificity and sensitivity in 
      three human cell lines. The designed substrates represent the first-in-a class of 
      new substrates that can be utilized to screen hHEXA inhibitors in adherent human 
      cultured cells.
FAU - Miura, Kazuki
AU  - Miura K
AD  - Department of Chemistry and Life Science, College of Bioresource Sciences, Nihon 
      University.
FAU - Aoyama, Yuka
AU  - Aoyama Y
AD  - Department of Chemistry and Life Science, College of Bioresource Sciences, Nihon 
      University.
FAU - Natsu, Yurika
AU  - Natsu Y
AD  - Department of Chemistry and Life Science, College of Bioresource Sciences, Nihon 
      University.
FAU - Koyama, Ryosuke
AU  - Koyama R
AD  - Department of Chemistry and Life Science, College of Bioresource Sciences, Nihon 
      University.
FAU - Hirano, Takako
AU  - Hirano T
AD  - Department of Chemistry and Life Science, College of Bioresource Sciences, Nihon 
      University.
FAU - Nishio, Toshiyuki
AU  - Nishio T
AD  - Department of Chemistry and Life Science, College of Bioresource Sciences, Nihon 
      University.
FAU - Hakamata, Wataru
AU  - Hakamata W
AD  - Department of Chemistry and Life Science, College of Bioresource Sciences, Nihon 
      University.
LA  - eng
PT  - Journal Article
PL  - Japan
TA  - Chem Pharm Bull (Tokyo)
JT  - Chemical & pharmaceutical bulletin
JID - 0377775
RN  - 0 (Fluorescent Dyes)
RN  - EC 3.2.1.52 (beta-N-Acetylhexosaminidases)
SB  - IM
MH  - Fluorescent Dyes/chemical synthesis/*chemistry
MH  - HeLa Cells
MH  - Humans
MH  - Models, Molecular
MH  - Molecular Structure
MH  - *Optical Imaging
MH  - beta-N-Acetylhexosaminidases/*analysis/metabolism
OTO - NOTNLM
OT  - fluorogenic substrate
OT  - inhibitor screening
OT  - lysosomal storage disease
OT  - pharmacological chaperone
OT  - beta-N-acetyl-D-hexosaminidase
EDAT- 2020/06/02 06:00
MHDA- 2021/01/12 06:00
CRDT- 2020/06/02 06:00
PHST- 2020/06/02 06:00 [entrez]
PHST- 2020/06/02 06:00 [pubmed]
PHST- 2021/01/12 06:00 [medline]
AID - 10.1248/cpb.c20-00069 [doi]
PST - ppublish
SO  - Chem Pharm Bull (Tokyo). 2020;68(6):526-533. doi: 10.1248/cpb.c20-00069.