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TEPP-46-Based AIE Fluorescent Probe for Detection and Bioimaging of PKM2 in Living Cells
Dong Wang1, Chunmeng Li2, Ya Zhu2
1State Key Laboratory of Natural Medicines, Key Lab of Drug Metabolism and Pharmacokinetics, China Pharmaceutical University, 210009 Nanjing, China.
Abstract:
Pyruvate kinase (PK) M2 (PKM2), a glycolytic enzyme, is a hallmark of different types of tumors and plays a significant role in the Warburg effect. However, there is no fluorescent probe for PKM2 that has been reported yet. In this study, TEPC466, a novel TEPP-46-based aggregation-induced emission (AIE) probe for the detection of PKM2, was designed, synthesized, and fully characterized by 1H NMR, 13C NMR, and high-resolution mass spectrometry. When the fluorescent agent, coumarine, was conjugated to TEPP-46, the bioprobe TEPC466 showed a high degree of selectivity and sensitivity for the detection of PKM2 protein via the AIE effect. TEPC466 was then successfully applied in imaging the PKM2 protein in colorectal cancer cells with low toxicity. Moreover, structure-based modeling and the PK activity assay confirmed that TEPC466 has a better binding with PKM2 than TEPP-46, which suggests that TEPC466 could also be a good agonist of PKM2. Taken together, the bioprobe shows potential in selective detection of PKM2 and provides a useful tool for cancer diagnosis and therapy.
Insights
Researchers developed a novel fluorescent probe, TEPC466, for detecting pyruvate kinase M2 (PKM2), an enzyme crucial in cancer. This probe enables sensitive imaging of PKM2 in colorectal cancer cells, offering potential for cancer diagnosis and therapy.
Area of Science:
- Biochemistry
- Molecular Biology
- Cancer Research
Background:
- Pyruvate kinase M2 (PKM2) is a key glycolytic enzyme implicated in various cancers and the Warburg effect.
- A specific fluorescent probe for PKM2 detection has been lacking, hindering its study.
Purpose of the Study:
- To design, synthesize, and characterize a novel aggregation-induced emission (AIE) fluorescent probe for selective PKM2 detection.
- To evaluate the probe's efficacy in imaging PKM2 in cancer cells and assess its potential as a PKM2 agonist.
Main Methods:
- Design and synthesis of the TEPP-46 based AIE probe, TEPC466, by conjugating coumarine.
- Characterization using 1H NMR, 13C NMR, and high-resolution mass spectrometry.
- In vitro assays for selectivity, sensitivity, PKM2 imaging in colorectal cancer cells, and PK activity assays.
Main Results:
- TEPC466 demonstrated high selectivity and sensitivity for PKM2 detection through the AIE effect.
- Successful imaging of PKM2 protein in colorectal cancer cells with low toxicity.
- TEPC466 showed enhanced binding to PKM2 compared to TEPP-46, suggesting agonistic potential.
Conclusions:
- The novel AIE probe TEPC466 enables selective and sensitive detection of PKM2.
- TEPC466 is a valuable tool for visualizing PKM2 in cancer cells and shows promise for cancer diagnosis and therapeutic strategies.
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