An Economical High‐Throughput “FP‐Tag” Assay for Screening Glycosyltransferase Inhibitors**

Yin, Xinjian; Li, Jiaxin; Chen, Senhua; Wu, Yuping; She, Zhigang; Liu, Lan; Wang, Yue; Gao, Zhizeng

doi:10.1002/cbic.202000746

Cited by 4 publications

(3 citation statements)

References 43 publications

(16 reference statements)

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“…As a marine natural product research group, we have a long-term interest to find bioactive molecules from the marine environment, and indeed we have previously successfully identified several potent inhibitors from marine natural product libraries with FP-based assays [31] , [32] . Therefore, we performed a pilot HTS with a small marine natural product library (about 300 compounds) at 10 µM ( Figure 7 A).…”

Section: Resultsmentioning

confidence: 99%

A robust high-throughput fluorescent polarization assay for the evaluation and screening of SARS-CoV-2 fusion inhibitors

Yin

Chen

Yuan

et al. 2021

Bioorganic Chemistry

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Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) is a serious threat to global health. One attractive antiviral target is the membrane fusion mechanism employed by the virus to gain access to the host cell. Here we report a robust protein-based fluorescent polarization assay, that mimicking the formation of the six-helix bundle (6-HB) process during the membrane fusion, for the evaluation and screening of SARS-CoV-2 fusion Inhibitors. The IC 50 of known inhibitors, HR2P, EK1, and Salvianolic acid C (Sal C) were measured to be 6 nM, 2.5 nM, and 8.9 µM respectively. In addition, we found Sal A has a slightly lower IC 50 (3.9 µM) than Sal C. Interestingly, simple caffeic acid can also disrupt the formation of 6-HB with a sub-mM concentration. Pilot high throughput screening (HTS) of a small marine natural product library validates the assay with a Z’ factor close to 0.8. We envision the current assay provides a convenient way to screen SARS-CoV-2 fusion inhibitors and assess their binding affinity.

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Section: Resultsmentioning

confidence: 99%

A robust high-throughput fluorescent polarization assay for the evaluation and screening of SARS-CoV-2 fusion inhibitors

Yin

Chen

Yuan

et al. 2021

Bioorganic Chemistry

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show abstract

“…As a marine natural product research group, we have a long-term interest to find bioactive molecules from the marine environment, and indeed we have previously successfully identified several potent inhibitors from marine natural product libraries with FP-based assays [31-32] . Therefore, we performed a pilot HTS with a small marine natural product library (about 300 compounds) at 10 µM (Figure 7A).…”

Section: Resultsmentioning

confidence: 99%

A Robust High-throughput Fluorescent Polarization Assay for the Evaluation and Screening of SARS-CoV-2 Fusion Inhibitors

Yin

Chen

Yuan

et al. 2021

Preprint

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Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) is a serious threat to global health. One attractive antiviral target is the membrane fusion mechanism employed by the virus to gain access to the host cell. Here we report a robust protein-based fluorescent polarization assay, that mimicking the formation of the six-helix bundle (6-HB) process during the membrane fusion, for the evaluation and screening of SARS-CoV-2 fusion Inhibitors. The IC50 of known inhibitors, HR2P, EK1, and Salvianolic acid C (Sal C) were measured to be 6 nM, 2.5 nM, and 8.9 uM respectively. In addition, we found Sal A has a slightly lower IC50 (3.9 uM) than Sal C. Interesting, simple caffeic acid can also disrupt the formation of 6-HB with sub-mM concentration. A pilot high throughput screening (HTS) a small marine natural product library validates the assay with a Z factor close to 0.8. We envision the current assay provides a convenient way to screen SARS-CoV-2 fusion inhibitor and assess their binding affinity.

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“…Based on this method, 2-methyleurotinone was identified as a low-μM OGT inhibitor. The BSA-based "FP-Tag" will be applicable to other types of glycosyltransferases 49 . In 2020, Seebald et al reported the synthesis and application of fluorescent nucleoside analogues as the foundation for directed approaches for competitive binding analyses.…”

Section: Fluorescently Modified Glycosyl Donors and Acceptorsmentioning

confidence: 99%

Recent advances in rapid screening methods for glycosyltransferases

Xia,

Huang,

Zhang

et al. 2023

Preprint

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Glycosyltransferases (GTs) are a diverse group of transferases widely distributed in microorganisms and plants which catalyze the transfer of glycosyl moieties from activated sugar donors to a diverse range of acceptor molecules. GTs play important roles in biological processes, such as cell signal transduction, cell-cell interactions, immune response and diseases. In addition, the utilization of glycosyltransferases to synthesize glycosides has become a major area of pharmaceutical research. The construction of a rapid and efficient screening method for glycosyltransferases is the key to the successful discovery, identification, and modification of new enzymes. In the past few decades, a series of screening methods have been developed to assess activity. This review focuses on various methods for rapid screening of GTs, outlining their advantages and disadvantages.

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An Economical High‐Throughput “FP‐Tag” Assay for Screening Glycosyltransferase Inhibitors**

Cited by 4 publications

References 43 publications

A robust high-throughput fluorescent polarization assay for the evaluation and screening of SARS-CoV-2 fusion inhibitors

A robust high-throughput fluorescent polarization assay for the evaluation and screening of SARS-CoV-2 fusion inhibitors

A Robust High-throughput Fluorescent Polarization Assay for the Evaluation and Screening of SARS-CoV-2 Fusion Inhibitors

Recent advances in rapid screening methods for glycosyltransferases

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