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Nano Research 2023, 16(1): 894-904 https://doi.org/10.1007/s12274-022-4785-5
Research Article | Issue | Published: 02 September 2022

Probing cell membrane integrity using a histone-targeting protein nanocage displaying precisely positioned fluorophores

Show Author's Information Ti Fang1,2,  Chaoqun Li2,5,  Ao Liang2,5,  Hui Zhang2,  Fan Zhang2,  Xian-En Zhang3,4,5,  Yi-Yu Yang1( ),  Feng Li2,5( )
Guangzhou Women and Children’s Medical Center, Guangzhou Medical University, Guangzhou 510120, China
State Key Laboratory of Virology, Wuhan Institute of Virology, Center for Biosafety Mega-Science, Chinese Academy of Sciences, Wuhan 430071, China
Faculty of Synthetic Biology, Shenzhen Institute of Advanced Technology, Chinese Academy of Sciences, Shenzhen 518055, China
National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China
University of Chinese Academy of Sciences, Beijing 100049, China
Keywords:
nanoparticles, fluorescent probes, protein engineering, cell membrane damage, ferritin superfamily
Topics:
BiotechnologyCell membranes Nanoprobes
Cite this article:
Fang T, Li C, Liang A, et al. Probing cell membrane integrity using a histone-targeting protein nanocage displaying precisely positioned fluorophores. Nano Research, 2023, 16(1): 894-904. https://doi.org/10.1007/s12274-022-4785-5
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Abstract Full text Electronic supplementary material About this article

Cell membrane integrity is fundamental to the normal activities of cells and is involved in both acute and chronic pathologies. Here, we report a probe for analyzing cell membrane integrity developed from a 9 nm-sized protein nanocage named Dps via fluorophore conjugation with high spatial precision to avoid self-quenching. The probe cannot enter normal live cells but can accumulate in dead or live cells with damaged membranes, which, interestingly, leads to weak cytoplasmic and strong nuclear staining. This differential staining is found attributed to the high affinity of Dps for histones rather than DNA, providing a staining mechanism different from those of known membrane exclusion probes (MEPs). Moreover, the Dps nanoprobe is larger in size and thus applies a more stringent criterion for identifying severe membrane damage than currently available MEPs. This study shows the potential of Dps as a new bioimaging platform for biological and medical analyses.


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Electronic supplementary material
About this article

Probing cell membrane integrity using a histone-targeting protein nanocage displaying precisely positioned fluorophores

Show Author's information Ti Fang1,2, Chaoqun Li2,5, Ao Liang2,5, Hui Zhang2, Fan Zhang2, Xian-En Zhang3,4,5, Yi-Yu Yang1( ), Feng Li2,5( )
Guangzhou Women and Children’s Medical Center, Guangzhou Medical University, Guangzhou 510120, China
State Key Laboratory of Virology, Wuhan Institute of Virology, Center for Biosafety Mega-Science, Chinese Academy of Sciences, Wuhan 430071, China
Faculty of Synthetic Biology, Shenzhen Institute of Advanced Technology, Chinese Academy of Sciences, Shenzhen 518055, China
National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China
University of Chinese Academy of Sciences, Beijing 100049, China

Abstract

Cell membrane integrity is fundamental to the normal activities of cells and is involved in both acute and chronic pathologies. Here, we report a probe for analyzing cell membrane integrity developed from a 9 nm-sized protein nanocage named Dps via fluorophore conjugation with high spatial precision to avoid self-quenching. The probe cannot enter normal live cells but can accumulate in dead or live cells with damaged membranes, which, interestingly, leads to weak cytoplasmic and strong nuclear staining. This differential staining is found attributed to the high affinity of Dps for histones rather than DNA, providing a staining mechanism different from those of known membrane exclusion probes (MEPs). Moreover, the Dps nanoprobe is larger in size and thus applies a more stringent criterion for identifying severe membrane damage than currently available MEPs. This study shows the potential of Dps as a new bioimaging platform for biological and medical analyses.

Keywords: nanoparticles, fluorescent probes, protein engineering, cell membrane damage, ferritin superfamily

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Publication history
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Acknowledgements

Publication history

Received: 07 May 2022
Revised: 13 July 2022
Accepted: 18 July 2022
Published: 02 September 2022
Issue date: January 2023

Copyright

© Tsinghua University Press 2022

Acknowledgements

This work was financially supported by the National Natural Science Foundation of China (No. 31771103), Chinese Academy of Sciences (CAS) Emergency Project of African Swine Fever (ASF) Research (No. KJZD-SW-L07), and the Scientific Instrument Developing Project of the CAS (No. YJKYYQ20190057). We thank Juan Min and Ding Gao at the Core Facility and Technical Support of Wuhan Institute of Virology for assistance in tissue imaging, confocal microscopy, and transmission electron microscopy.

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