DNA-based probes for flow cytometry analysis of endocytosis and recycling

Claire Fanny Marie Dumont, Ewa Czuba, Moore Chen, Jose A Villadangos, Angus P.R. Johnston, Justine D. Mintern

Research output: Contribution to journalArticleResearchpeer-review

1 Citation (Scopus)

Abstract

The internalization of proteins plays a key role in cell development, cell signaling and immunity. We have previously developed a specific hybridization internalization probe (SHIP) to quantitate the internalization of proteins and particles into cells. Herein, we extend the utility of SHIP to examine both the endocytosis and recycling of surface receptors using flow cytometry. SHIP was used to monitor endocytosis of membrane-bound transferrin receptor (TFR) and its soluble ligand transferrin (TF). SHIP enabled measurements of the proportion of surface molecules internalized, the internalization kinetics and the proportion and rate of internalized molecules that recycle to the cell surface with time. Using this method, we have demonstrated the internalization and recycling of holo-TF and an antibody against the TFR behave differently. This assay therefore highlights the implications of receptor internalization and recycling, where the internalization of the receptor-antibody complex behaves differently to the receptor-ligand complex. In addition, we observe distinct internalization patterns for these molecules expressed by different subpopulations of primary cells. SHIP provides a convenient and high throughput technique for analysis of trafficking parameters for both cell surface receptors and their ligands.

Original languageEnglish
Pages (from-to)242-249
Number of pages8
JournalTraffic
Volume18
Issue number4
DOIs
Publication statusPublished - 1 Apr 2017

Keywords

  • endocytosis
  • probes
  • recycling
  • transferrin

Cite this

Dumont, C. F. M., Czuba, E., Chen, M., Villadangos, J. A., Johnston, A. P. R., & Mintern, J. D. (2017). DNA-based probes for flow cytometry analysis of endocytosis and recycling. Traffic, 18(4), 242-249. https://doi.org/10.1111/tra.12466
Dumont, Claire Fanny Marie ; Czuba, Ewa ; Chen, Moore ; Villadangos, Jose A ; Johnston, Angus P.R. ; Mintern, Justine D. / DNA-based probes for flow cytometry analysis of endocytosis and recycling. In: Traffic. 2017 ; Vol. 18, No. 4. pp. 242-249.
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Dumont, CFM, Czuba, E, Chen, M, Villadangos, JA, Johnston, APR & Mintern, JD 2017, 'DNA-based probes for flow cytometry analysis of endocytosis and recycling', Traffic, vol. 18, no. 4, pp. 242-249. https://doi.org/10.1111/tra.12466

DNA-based probes for flow cytometry analysis of endocytosis and recycling. / Dumont, Claire Fanny Marie; Czuba, Ewa; Chen, Moore; Villadangos, Jose A; Johnston, Angus P.R.; Mintern, Justine D.

In: Traffic, Vol. 18, No. 4, 01.04.2017, p. 242-249.

Research output: Contribution to journalArticleResearchpeer-review

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AU - Mintern, Justine D.

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Dumont CFM, Czuba E, Chen M, Villadangos JA, Johnston APR, Mintern JD. DNA-based probes for flow cytometry analysis of endocytosis and recycling. Traffic. 2017 Apr 1;18(4):242-249. https://doi.org/10.1111/tra.12466