Young Researcher Paper Award 2023

Notice of retraction
Vol. 34, No. 8(3), S&M3042

Notice of retraction
Vol. 32, No. 8(2), S&M2292

Print: ISSN 0914-4935
Online: ISSN 2435-0869
Sensors and Materials
is an international peer-reviewed open access journal to provide a forum for researchers working in multidisciplinary fields of sensing technology.
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Orthogonality of α-Sulfoquinovosidase in Human Cells and Development of Its Fluorescent Substrate

Ryosuke Yoshida, Ryosei Kaguma, Ryosuke Kaneko, Ichiro Matuso, Makoto Yoritate, Go Hirai, Takamasa Teramoto, Yoshimitsu Kakuta, Kosuke Minamihata, Noriho Kamiya, Teruki Nii, Akihiro Kishimura, Takeshi Mori, and Yoshiki Katayama

(Received December 4, 2023; Accepted March 28, 2024)

Keywords: enzyme, cell ELISA, fluorescence

Human orthogonal enzymes (HOEs), do not show the same activities as the endogenous enzymes of human cells and thus are useful as amplification enzymes to detect antigen proteins in biological samples. Here, we evaluates a new HOE from Escherichia coli, α-sulfoquinovosidase (α-SQase). We confirmed that the activity of α-SQase did not exist in examined human cell lines, and thus it was applicable to live-cell enzyme-linked immunosorbent assay (ELISA) in which the antigen membrane protein on cells was detected without inactivating endogenous enzymes, a pretreatment required for cell ELISA using conventional amplification enzymes. Here, we also developed a fluorescent substrate for α-SQase whose active residue is located at the end of the narrow, deep pocket of the substrate recognition site. The designed methylumbelliferyl substrate with a hydroxyl benzyl alcohol linker showed similar reactivity as the p-nitrophenol substrate, a good substrate for α-SQase.

Corresponding author: Takeshi Mori and Yoshiki Katayama

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