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Related Concept Videos

Lysosomes01:31

Lysosomes

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Lysosomes are membrane-enclosed spherical sacs derived from the Golgi apparatus. The most important function of the lysosome is degrading macromolecules and biological polymers that are released during membrane trafficking events such as the secretory, endocytic, autophagic, and phagocytic pathways. The degradation is carried out by several hydrolytic enzymes active in an acidic environment of the lysosomal lumen. These acid hydrolases are involved in cellular processes such as cell signaling,...
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Receptor-mediated Endocytosis01:39

Receptor-mediated Endocytosis

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Overview
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Receptor-mediated Endocytosis01:20

Receptor-mediated Endocytosis

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Receptor-mediated endocytosis is when bulk amounts of specific molecules are imported into a cell after binding to cell surface receptors. The molecules bound to these receptors are taken into the cell through inward folding of the cell surface membrane, which is eventually pinched off into a vesicle within the cell. Structural proteins, such as clathrin, coat the budding vesicle.
Clathrin-Mediated Endocytosis of LDL
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Delivery Pathways to the Lysosome01:36

Delivery Pathways to the Lysosome

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Eukaryotic cells use different mechanisms to eliminate toxic waste obsolete and worn-out substances. Lysosomes play a pivotal role in this, and hence, these substances are carried to the lysosome from other parts of the cell and extracellular space through different pathways. The most elaborately studied pathways to the lysosome are the endocytic pathways.
Endocytosis
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Lysosomal Hydrolases01:22

Lysosomal Hydrolases

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Lysosomes are the site for the degradation of macromolecules and biological polymers released during membrane trafficking events such as secretory, endocytic, autophagic, and phagocytic pathways. The membrane-enclosed area of the lysosome, called the lumen, contains hydrolytic enzymes active in an acidic environment. These acid hydrolases are functional at a pH between 4.5 and 5 and are involved in cellular processes such as cell signaling, energy metabolism, restoration of the plasma membrane,...
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Clathrin Coated Vesicles01:12

Clathrin Coated Vesicles

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Clathrin-coated vesicles use endocytosis to transport receptors and lysosomal hydrolases from the Golgi to the lysosome in the late secretory pathway. Clathrin-mediated endocytosis was the first described endocytic process, and Clathrin-coated vesicles remain one of the most well-studied transport vesicles. The molecular machinery that generates clathrin-coated vesicles comprises over 50 proteins that precisely coordinate vesicle formation. Cell surface receptors concentrated in indented sites...
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Related Experiment Video

Updated: Jan 10, 2026

Engineering Cell-permeable Protein
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Engineered calcium-regulated affinity protein for efficient internalization and lysosomal toxin delivery.

Malin Jönsson1, Marit Möller1, Leon Schierholz2

  • 1Department of Protein Science, SciLifeLab, KTH-Royal Institute of Technology, Stockholm, Sweden.

Proceedings of the National Academy of Sciences of the United States of America
|November 25, 2025
PubMed
Summary

Researchers developed a novel calcium-regulated protein binder for targeted cancer therapy. This engineered protein targets cancer cells, delivering toxins to lysosomes for potent cell killing, enhancing drug delivery efficacy.

Keywords:
calcium-regulated affinitycancerconditional targetingdrug-conjugateendosomal release

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Cancer Therapy

Background:

  • Protein-drug conjugates (PDCs) offer targeted cancer therapy by delivering cytotoxic payloads to tumor cells.
  • Effective PDC therapy relies on payload internalization and retention within target cells.
  • Current strategies face challenges in controlling payload release and receptor recycling.

Purpose of the Study:

  • To engineer a protein domain with calcium-regulated affinity for controlled target binding and lysosomal trafficking.
  • To develop a novel drug delivery system for targeted cancer treatment.
  • To assess the efficacy of the calcium-regulated binder in delivering toxins to cancer cells.

Main Methods:

  • Engineering of an Epidermal Growth Factor Receptor (EGFR) binder with calcium-regulated affinity (CaRA).
  • Affinity measurements and structural modeling to understand calcium-modulated binding.
  • Live cell imaging to track internalization, lysosomal trafficking, and receptor fate.
  • Cytotoxicity assays to determine the efficacy of the toxin delivery system.

Main Results:

  • The CaRA_EGFR binder demonstrated calcium-dependent affinity, facilitating endosomal dissociation.
  • Live cell imaging confirmed efficient internalization and lysosomal trafficking of the binder, with EGFR recycling.
  • CaRA_EGFR effectively delivered toxins to lysosomes, achieving potent cytotoxicity (IC50 = 0.8 nM) in EGFR-expressing cancer cells.

Conclusions:

  • Engineered calcium-regulated protein domains can achieve targeted payload delivery and lysosomal trafficking independent of receptor fate.
  • This approach offers a novel strategy for enhancing the efficacy of protein-drug conjugates in cancer therapy.
  • CaRA_EGFR represents a promising tool for developing next-generation targeted cancer therapeutics.