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Tagging and Fusion Proteins

Proteins are involved in several cellular processes and biochemical reactions. Analyzing a specific protein of interest requires it to be isolated from the other proteins in the cell. This is achieved by overexpressing the specific gene in a suitable host to produce large quantities of the target protein. A tag or label is recombined with the gene to produce a fusion protein containing the target protein and the tag. The tags on these fusion proteins can then be used for easy detection and...
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Related Experiment Video

Updated: May 31, 2026

Delivery of Proteins, Peptides or Cell-impermeable Small Molecules into Live Cells by Incubation with the Endosomolytic Reagent dfTAT
10:30

Delivery of Proteins, Peptides or Cell-impermeable Small Molecules into Live Cells by Incubation with the Endosomolytic Reagent dfTAT

Published on: September 2, 2015

Dithiolane-Based Reversible "Trojan Tag" For Intracellular Protein Delivery and Prodrug Design.

Yahui He1, Xiaoman Yu1, Yuanhao Dai1

  • 1Beijing National Laboratory for Molecular Sciences, Center for Soft Matter Science and Engineering, Key Laboratory of Polymer Chemistry and Physics of the Ministry of Education, College of Chemistry and Molecular Engineering, Peking University, Beijing 100871, People's Republic of China.

ACS Applied Materials & Interfaces
|May 29, 2026
PubMed
Summary
This summary is machine-generated.

Researchers developed a dithiolane-based "Trojan tag" (DTL tag) for protein prodrugs. This tag enables efficient delivery, reversible activity switching, and traceless intracellular activation, enhancing cancer therapy.

Keywords:
dithiolaneintracellular deliveryprodrugprotein therapeuticstraceless release

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Area of Science:

  • Biochemistry
  • Chemical Biology
  • Drug Delivery

Background:

  • Prodrug strategies are crucial for controlling protein therapeutics' activity and delivery.
  • Current methods face challenges in achieving universal platforms for reversible activity switching and efficient cellular internalization.
  • Developing modular systems for protein prodrugs is essential for targeted therapies.

Purpose of the Study:

  • To introduce a novel dithiolane-based "Trojan tag" (DTL tag) for constructing modular protein prodrugs.
  • To demonstrate the DTL tag's capability for efficient protein delivery and intracellular activation.
  • To showcase the DTL tag's potential in enhancing therapeutic protein efficacy.

Main Methods:

  • Conjugation of the DTL tag to various proteins, including an anti-PD-L1 nanobody and enzymes like luciferase and RNase A.
  • Assessment of cellular internalization efficiency of DTL-tagged proteins.
  • Evaluation of reduction-triggered activity switching and intracellular reactivation of enzymes.
  • Combination with transfection reagents to study cytosolic delivery and therapeutic effects.

Main Results:

  • The DTL tag facilitated efficient cellular internalization of diverse proteins, irrespective of their size and isoelectric point.
  • Application to an anti-PD-L1 nanobody resulted in enhanced internalization and effective reduction of PD-L1 on tumor cells.
  • The DTL tag acted as a reduction-triggered switch, enabling reactivation of enzyme activity post-delivery.
  • Synergistic effects with transfection reagents improved cytosolic delivery of RNase A, leading to enhanced cancer cell killing.

Conclusions:

  • The DTL tag provides a simple, multifunctional platform for creating modular protein prodrugs with traceless intracellular activation.
  • This strategy offers a new approach for developing smart protein therapeutics with controlled activity and improved delivery.
  • The DTL tag platform streamlines the development of protein prodrugs for enhanced therapeutic applications.