A Universal and Programmable Platform based on Fluorescent Peptide-Conjugated Probes for Detection of Proteins in Organelles of Living Cells
Dr. Xia Wu
State Key Laboratory of Biogeology and Environmental Geology, Faculty of Materials Science and Chemistry, China University of Geosciences, 430074 Wuhan, China
These authors contributed equally to this work.
Contribution: Conceptualization (lead), Data curation (lead), Formal analysis (lead), Funding acquisition (supporting), Writing - original draft (lead)
Search for more papers by this authorDr. Jing-Jing Hu
State Key Laboratory of Biogeology and Environmental Geology, Faculty of Materials Science and Chemistry, China University of Geosciences, 430074 Wuhan, China
These authors contributed equally to this work.
Contribution: Conceptualization (supporting), Funding acquisition (equal), Writing - review & editing (lead)
Search for more papers by this authorDr. Chong Duan
State Key Laboratory of Biogeology and Environmental Geology, Faculty of Materials Science and Chemistry, China University of Geosciences, 430074 Wuhan, China
Contribution: Data curation (supporting), Formal analysis (supporting)
Search for more papers by this authorDr. Rui Liu
State Key Laboratory of Biogeology and Environmental Geology, Faculty of Materials Science and Chemistry, China University of Geosciences, 430074 Wuhan, China
Contribution: Data curation (supporting)
Search for more papers by this authorProf. Fan Xia
State Key Laboratory of Biogeology and Environmental Geology, Faculty of Materials Science and Chemistry, China University of Geosciences, 430074 Wuhan, China
Contribution: Conceptualization (lead), Funding acquisition (lead), Supervision (lead), Writing - review & editing (lead)
Search for more papers by this authorCorresponding Author
Prof. Xiaoding Lou
State Key Laboratory of Biogeology and Environmental Geology, Faculty of Materials Science and Chemistry, China University of Geosciences, 430074 Wuhan, China
Contribution: Conceptualization (lead), Funding acquisition (lead), Supervision (equal), Writing - review & editing (equal)
Search for more papers by this authorDr. Xia Wu
State Key Laboratory of Biogeology and Environmental Geology, Faculty of Materials Science and Chemistry, China University of Geosciences, 430074 Wuhan, China
These authors contributed equally to this work.
Contribution: Conceptualization (lead), Data curation (lead), Formal analysis (lead), Funding acquisition (supporting), Writing - original draft (lead)
Search for more papers by this authorDr. Jing-Jing Hu
State Key Laboratory of Biogeology and Environmental Geology, Faculty of Materials Science and Chemistry, China University of Geosciences, 430074 Wuhan, China
These authors contributed equally to this work.
Contribution: Conceptualization (supporting), Funding acquisition (equal), Writing - review & editing (lead)
Search for more papers by this authorDr. Chong Duan
State Key Laboratory of Biogeology and Environmental Geology, Faculty of Materials Science and Chemistry, China University of Geosciences, 430074 Wuhan, China
Contribution: Data curation (supporting), Formal analysis (supporting)
Search for more papers by this authorDr. Rui Liu
State Key Laboratory of Biogeology and Environmental Geology, Faculty of Materials Science and Chemistry, China University of Geosciences, 430074 Wuhan, China
Contribution: Data curation (supporting)
Search for more papers by this authorProf. Fan Xia
State Key Laboratory of Biogeology and Environmental Geology, Faculty of Materials Science and Chemistry, China University of Geosciences, 430074 Wuhan, China
Contribution: Conceptualization (lead), Funding acquisition (lead), Supervision (lead), Writing - review & editing (lead)
Search for more papers by this authorCorresponding Author
Prof. Xiaoding Lou
State Key Laboratory of Biogeology and Environmental Geology, Faculty of Materials Science and Chemistry, China University of Geosciences, 430074 Wuhan, China
Contribution: Conceptualization (lead), Funding acquisition (lead), Supervision (equal), Writing - review & editing (equal)
Search for more papers by this authorAbstract
Realizing protein analysis in organelles of living cells is of great significance for developing diagnostic and therapeutic methods of diseases. Fluorescent-labeled antibodies with well imaging performance and high affinity are classical biochemical tools for protein analysis, while due to the inability to effectively enter into cells, not to mention organelles and the uncontrollable reaction sites that might cause antibodies inactivation when chemically modification, they are hard to apply to living cells. Inspired by the structure of fluorescent-labeled antibodies, we designed as a universal detection platform that was based on the peptide-conjugated probes (PCPs) and consisted of three parts: a) a rotor type fluorescent molecular scaffold for conjugation and signal output; b) the cell penetration protein recognition unit; c) the subcellular organelle targeting unit. In living cells, PCPs could firstly localize at organelles and then proceed protein specific recognition, thus jointly leading to the restriction of twisted intramolecular charge transfer and activation of fluorescence signal. As a proof-of-concept, six different proteins in three typical intracellular organelles could be detected by our platform through simply replacing the recognition sequence of proteins and matching organelle targeting units. The position and intensity of fluorescence signals demonstrated specificity of PCPs and universality of the platform.
Conflict of interests
The authors declare no conflict of interest.
Open Research
Data Availability Statement
The data that support the findings of this study are available in the supplementary material of this article.
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