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Novel gene-encoded intermolecular FRET sensor for tracking glycosylation of CD147 in living cells

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Analytical and Bioanalytical Chemistry Aims and scope Submit manuscript

Abstract

CD147 is involved in various physiological processes and plays important roles for tumor metastasis. Glycosylation of the protein determines numerous functions of CD147. Up to now, hardly any sensor has been developed for detecting glycosylation of CD147 in live cells. There is a pressing requirement of development of a selective and continuous biosensor for cell imaging. The emergence of gene-encoded fluorescence resonance energy transfer (FRET) sensor provides a new way to develop the sensors to analysts. We designed and constructed novel gene-encoded FRET proteins sensing glycosylation of CD147 by measuring FRET ratio of two intermolecular motifs. With the decrease of CD147 glycosylation level in cells, the FRET ratio increased significantly. The specificity of the sensor targeting to CD147 was also determined by siRNA interference experiment. Finally, continuous living cell image of deglycosylation process of CD147 using the newly developed sensor has been performed successfully. The work not only provides useful tools for analyzing glycosylation of CD147 in living cells, but also implicates alternative strategy for detecting other glycosylated proteins.

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Funding

This research was funded by Major Research Plan of the National Natural Science Foundation of China(2016YFA0101200), National Natural Science Foundation of China (Projects 21778050, 91749125), Youth Fund of National Natural Science Foundation of China (81701232), New Medical Science Joint Fund of USTC (WK2070000123), and Training Program of the Major Foundation of USTC (WK3520000007).

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Author notes

  1. Hui Lu and Lingling Zhu contributed equally to this work.

Authors and Affiliations

  1. Hefei National Laboratory for Physical Sciences at Microscale, the CAS Key Laboratory of Innate Immunity and Chronic Disease, School of Basic Medical Sciences, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, 230027, Anhui, China

    Hui Lu, Lingling Zhu, Wenqian Li, Yushu Ge & Dan Liu

  2. Abbisko Therapeutics Co., Ltd., Shanghai, 200120, China

    Shuangyun Hu

  3. The First Affiliated Hospital of University of Science and Technology of China, Hefei, 230001, Anhui, China

    Dan Liu

Authors
  1. Hui Lu
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  2. Lingling Zhu
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  3. Wenqian Li
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  4. Shuangyun Hu
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  5. Yushu Ge
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  6. Dan Liu
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Correspondence to Yushu Ge or Dan Liu.

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Lu, H., Zhu, L., Li, W. et al. Novel gene-encoded intermolecular FRET sensor for tracking glycosylation of CD147 in living cells. Anal Bioanal Chem 413, 3193–3199 (2021). https://doi.org/10.1007/s00216-021-03256-0

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  • DOI: https://doi.org/10.1007/s00216-021-03256-0

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