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Fluid-Phase Endocytosis and Lysosomal Degradation of Bovine Lactoferrin in Lung Cells
The iron-binding protein lactoferrin and the cell-penetrating peptides derived from its sequence utilise endocytosis to enter different cell types. The full-length protein has been extensively investigated as a potential therapeutic against a range of pathogenic bacteria, fungi, and viruses, includi...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9032238/ https://www.ncbi.nlm.nih.gov/pubmed/35456688 http://dx.doi.org/10.3390/pharmaceutics14040855 |
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author | Sayers, Edward John Palmer, Iwan Hope, Lucy Hope, Paul Watson, Peter Jones, Arwyn Tomos |
author_facet | Sayers, Edward John Palmer, Iwan Hope, Lucy Hope, Paul Watson, Peter Jones, Arwyn Tomos |
author_sort | Sayers, Edward John |
collection | PubMed |
description | The iron-binding protein lactoferrin and the cell-penetrating peptides derived from its sequence utilise endocytosis to enter different cell types. The full-length protein has been extensively investigated as a potential therapeutic against a range of pathogenic bacteria, fungi, and viruses, including SARS-CoV-2. As a respiratory antiviral agent, several activity mechanisms have been demonstrated for lactoferrin, at the extracellular and plasma membrane levels, but as a protein that enters cells it may also have intracellular antiviral activity. Characterisation of lactoferrin’s binding, endocytic traffic to lysosomes, or recycling endosomes for exocytosis is lacking, especially in lung cell models. Here, we use confocal microscopy, flow cytometry, and degradation assays to evaluate binding, internalisation, endocytic trafficking, and the intracellular fate of bovine lactoferrin in human lung A549 cells. In comparative studies with endocytic probes transferrin and dextran, we show that lactoferrin binds to negative charges on the cell surface and actively enters cells via fluid-phase endocytosis, in a receptor-independent manner. Once inside the cell, we show that it is trafficked to lysosomes where it undergoes degradation within two hours. These findings provide opportunities for investigating both lactoferrin and derived cell-penetrating peptides activities of targeting intracellular pathogens. |
format | Online Article Text |
id | pubmed-9032238 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-90322382022-04-23 Fluid-Phase Endocytosis and Lysosomal Degradation of Bovine Lactoferrin in Lung Cells Sayers, Edward John Palmer, Iwan Hope, Lucy Hope, Paul Watson, Peter Jones, Arwyn Tomos Pharmaceutics Article The iron-binding protein lactoferrin and the cell-penetrating peptides derived from its sequence utilise endocytosis to enter different cell types. The full-length protein has been extensively investigated as a potential therapeutic against a range of pathogenic bacteria, fungi, and viruses, including SARS-CoV-2. As a respiratory antiviral agent, several activity mechanisms have been demonstrated for lactoferrin, at the extracellular and plasma membrane levels, but as a protein that enters cells it may also have intracellular antiviral activity. Characterisation of lactoferrin’s binding, endocytic traffic to lysosomes, or recycling endosomes for exocytosis is lacking, especially in lung cell models. Here, we use confocal microscopy, flow cytometry, and degradation assays to evaluate binding, internalisation, endocytic trafficking, and the intracellular fate of bovine lactoferrin in human lung A549 cells. In comparative studies with endocytic probes transferrin and dextran, we show that lactoferrin binds to negative charges on the cell surface and actively enters cells via fluid-phase endocytosis, in a receptor-independent manner. Once inside the cell, we show that it is trafficked to lysosomes where it undergoes degradation within two hours. These findings provide opportunities for investigating both lactoferrin and derived cell-penetrating peptides activities of targeting intracellular pathogens. MDPI 2022-04-13 /pmc/articles/PMC9032238/ /pubmed/35456688 http://dx.doi.org/10.3390/pharmaceutics14040855 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Sayers, Edward John Palmer, Iwan Hope, Lucy Hope, Paul Watson, Peter Jones, Arwyn Tomos Fluid-Phase Endocytosis and Lysosomal Degradation of Bovine Lactoferrin in Lung Cells |
title | Fluid-Phase Endocytosis and Lysosomal Degradation of Bovine Lactoferrin in Lung Cells |
title_full | Fluid-Phase Endocytosis and Lysosomal Degradation of Bovine Lactoferrin in Lung Cells |
title_fullStr | Fluid-Phase Endocytosis and Lysosomal Degradation of Bovine Lactoferrin in Lung Cells |
title_full_unstemmed | Fluid-Phase Endocytosis and Lysosomal Degradation of Bovine Lactoferrin in Lung Cells |
title_short | Fluid-Phase Endocytosis and Lysosomal Degradation of Bovine Lactoferrin in Lung Cells |
title_sort | fluid-phase endocytosis and lysosomal degradation of bovine lactoferrin in lung cells |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9032238/ https://www.ncbi.nlm.nih.gov/pubmed/35456688 http://dx.doi.org/10.3390/pharmaceutics14040855 |
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