ArticleTraffic (Copenhagen, Denmark)2020
An internally eGFP-tagged α-adaptin is a fully functional and improved fiduciary marker for clathrin-coated pit dynamics.
Article in Traffic (Copenhagen, Denmark), 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 12 papers.
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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
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Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
12 citing papers in PubMed, 21 citations in OpenAlex.
- Article
- Dynamic early recruitment of GAK-Hsc70 regulates coated pit maturation.Proceedings of the National Academy of Sciences of the United States of America · 2025Article
- Clathrin-associated carriers enable recycling through a kiss-and-run mechanism.Nature cell biology · 2024Article
- Far-Red Fluorescent Proteins: Tools for Advancing In Vivo Imaging.Biosensors · 2024Review
- FACI is a novel clathrin adaptor protein 2-binding protein that facilitates low-density lipoprotein endocytosis.Cell & bioscience · 2023Article
- Imaging nanoscale axial dynamics at the basal plasma membrane.The international journal of biochemistry & cell biology · 2023Review
- Resolution doubling in light-sheet microscopy via oblique plane structured illumination.Nature methods · 2022Article
- Increasing the field-of-view in oblique plane microscopy via optical tiling.Biomedical optics express · 2022Article
- FCHO controls AP2's initiating role in endocytosis through a PtdIns(4,5)PScience advances · 2022Article
- Imaging vesicle formation dynamics supports the flexible model of clathrin-mediated endocytosis.Nature communications · 2022Article
- CALM supports clathrin-coated vesicle completion upon membrane tension increase.Proceedings of the National Academy of Sciences of the United States of America · 2021Article
- An internally eGFP-tagged α-adaptin is a fully functional and improved fiduciary marker for clathrin-coated pit dynamics.Traffic (Copenhagen, Denmark) · 2020Article
Corrections and comments
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Authors and funding
4 authors at 1 institution in 1 country.
Funding
Abstract
Clathrin mediated endocytosis (CME) has been extensively studied in living cells by quantitative total internal reflection fluorescence microscopy (TIRFM). Fluorescent protein fusions to subunits of the major coat proteins, clathrin light chains or the heterotetrameric adaptor protein (AP2) complexes, have been used as fiduciary markers of clathrin coated pits (CCPs). However, the functionality of these fusion proteins has not been rigorously compared. Here, we generated stable cells lines overexpressing mRuby-CLCa and/or μ2-eGFP, σ2-eGFP, two markers currently in use, or a novel marker generated by inserting eGFP into the unstructured hinge region of the α subunit (α-eGFP). Using biochemical and TIRFM-based assays, we compared the functionality of the AP2 markers. All of the eGFP-tagged subunits were efficiently incorporated into AP2 and displayed greater accuracy in image-based CCP analyses than mRuby-CLCa. However, overexpression of either μ2-eGFP or σ2-eGFP impaired transferrin receptor uptake. In addition, μ2-eGFP reduced the rates of CCP initiation and σ2-eGFP perturbed AP2 incorporation into CCPs and CCP maturation. In contrast, CME and CCP dynamics were unperturbed in cells overexpressing α-eGFP. Moreover, α-eGFP was a more sensitive and accurate marker of CCP dynamics than mRuby-CLCa. Thus, our work establishes α-eGFP as a robust, fully functional marker for CME.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.