Qualitative and quantitative analysis of endocytic recycling

Reineke, James B.; Xie, Shuwei; Naslavsky, Naava; Caplan, Steve

doi:10.1016/bs.mcb.2015.04.002

Cited by 9 publications

(10 citation statements)

References 18 publications

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“…, protocol A). This approach has been used in several studies (Tanowitz and von Zastrow, ; Park et al, ; Vargas and Von Zastrow, ; Weber et al, ; Weigert et al, ; Barral et al, ; Millman et al, ; Henriques et al, ; Zuo et al, ; Finetti et al, ; Hsu et al, ; Reineke et al, ).…”

Section: Resultsmentioning

confidence: 99%

“…Thus, in this study, we performed analysis of protocols for quantitative assessment of endocytic recycling based on antibodies as a tool for convenient labeling of membrane proteins and monitoring their intra‐endosomal itinerary. Antibodies have been used for monitoring of endosomal recycling in a number of studies using various approaches and with diverse depth and fidelity of quantitative assessment (Ghosh et al, ; Tanowitz and Zastrow, 2003; Lin et al, ; Park et al, ; Scott et al, ; Vargas and Von Zastrow, ; Weber et al, ; Weigert et al, ; Chen et al, ; Jovanovic et al, ; Barral et al, ; Millman et al, ; Henriques et al, ; Ilić Tomaš et al, ; Mahmutefendić et al, ; Zuo et al, ; Zagorac et al, ; Finetti et al, ; Hsu et al, ; Reineke et al, ). In this report, we demonstrate that antibody‐based assays may be used for quantitative assessment of both clathrin‐dependent and clathrin‐independent membrane proteins, with the introduction of several upgrades that were required to fit the cellular physiology of endocytic trafficking and the inclusion of kinetic modeling.…”

Section: Discussionmentioning

confidence: 99%

“…In contrast to clathrin‐dependent cargo proteins, for many proteins that are endocytosed by the clathrin‐independent mechanism, the rate of endocytic uptake was not established and remains largely unknown. The best characterized is the recycling route of Major Histocompatibility Class I (MHC‐I) proteins (Weigert et al, ; Jovanovic et al, ; Donaldson and Williams, ; Grant and Donaldson, ), although recycling of several clathrin‐independent proteins was described in the last decade (Vargas and Von Zastrow, ; Weigert et al, ; Chen et al, ; Klein et al, ; Barral et al, ; Grant and Donaldson, ; Zuo et al, ; Maldonado‐Báez et al, ; Majeed et al, ; Hsu et al, ; Li et al, ; Reineke et al, ).…”

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confidence: 99%

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Quantitative Analysis of Endocytic Recycling of Membrane Proteins by Monoclonal Antibody-Based Recycling Assays

et al. 2016

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In this report, we present an analysis of several recycling protocols based on labeling of membrane proteins with specific monoclonal antibodies (mAbs). We analyzed recycling of membrane proteins that are internalized by clathrin-dependent endocytosis, represented by the transferrin receptor, and by clathrin-independent endocytosis, represented by the Major Histocompatibility Class I molecules. Cell surface membrane proteins were labeled with mAbs and recycling of mAb:protein complexes was determined by several approaches. Our study demonstrates that direct and indirect detection of recycled mAb:protein complexes at the cell surface underestimate the recycling pool, especially for clathrin-dependent membrane proteins that are rapidly reinternalized after recycling. Recycling protocols based on the capture of recycled mAb:protein complexes require the use of the Alexa Fluor 488 conjugated secondary antibodies or FITC-conjugated secondary antibodies in combination with inhibitors of endosomal acidification and degradation. Finally, protocols based on the capture of recycled proteins that are labeled with Alexa Fluor 488 conjugated primary antibodies and quenching of fluorescence by the anti-Alexa Fluor 488 displayed the same quantitative assessment of recycling as the antibody-capture protocols. J. Cell. Physiol. 232: 463-476, 2017. © 2016 Wiley Periodicals, Inc.

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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confidence: 99%

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Quantitative Analysis of Endocytic Recycling of Membrane Proteins by Monoclonal Antibody-Based Recycling Assays

et al. 2016

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“…In contrast to CDE cargo proteins, the recycling route was not established or is poorly characterized for many proteins that are endocytosed by the CIE. The best characterized is the recycling route of major histocompatibility class I (MHC-I) proteins [11], although recycling routes of several CIE cargo proteins have been described in the last decade [6,11,36].…”

Section: Endosomal Recyclingmentioning

confidence: 99%

Rapid Endosomal Recycling

Mahmutefendić¹,

Zagorac²,

Mačešić³

et al. 2018

Peripheral Membrane Proteins

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Peripheral membrane proteins are endocytosed by constitutive processes of membrane invaginations, followed by internalization driven by diverse endocytic machinery available at the cell surface. It is believed that after endocytic uptake, cargo proteins proceed either through the endosomal recycling circuit of the cell or travel toward late endosomes for degradation. In this chapter, we analyzed trafficking of seven cargo molecules (transferrin receptor, fully conformed MHC-I, non-conformed MHC-I, cholera-toxin B subunit, CD44, ICAM1, and G-protein-coupled receptor Rae-1) known to use the distinct endocytic route. For that purpose, we developed the software for multicompartment analysis of intracellular trafficking. We demonstrate that all endocytosed molecules are rapidly recycled and propose that the rapid recycling is a constitutive process that should be considered in the analysis of intracellular trafficking of peripheral membrane proteins. Keywords: rapid endosomal recycling, clathrin-independent cargo, endosomal recycling, endosomal trafficking, kinetic modeling of endosomal trafficking, transferrin receptor lular material, cell communication and information processing, motility, adhesion, and cell division (reviewed in [1-7]). Endocytic uptake occurs either by ligand binding to cell surface proteins (receptor-mediated endocytosis) or by uptake of extracellular fluid by membrane invaginations (fluid-phase endocytosis) [3]. Endocytosis is initiated by cellular proteins that change PM lipid composition and by the assembly of a series of cytoplasmic proteins,

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“…79, 80 The advent and widespread application of new imaging techniques such as super-resolution microscopy and quantitative methodology are also avenues that continue to contribute in our understanding of the molecular mechanism(s) involved in the processing of APP and tau and their roles in the pathogenesis of AD. 75, 81 …”

Section: Alzheimer's Disease and Evsmentioning

confidence: 99%

Emerging roles of extracellular vesicles in neurodegenerative disorders: focus on HIV-associated neurological complications

Lü

Cai

et al. 2016

Cell Death Dis

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Exosomes are membrane-enriched extracellular vesicles with a proposed diameter in the range of 30–100 nm. They are released during both normal homeostasis as well as under pathological conditions by most cell types. In recent years, there has been robust interest in the study of these vesicles as conduits for the delivery of information between cells in both analogous as well as disparate tissues. Their ability to transport specialized cargo including signaling mediators, proteins, messenger RNA and miRNAs characterizes these vesicles as primary facilitators of cell-to-cell communication and regulation. Exosomes have also been demonstrated to have important roles in the field of cancer biology and metastasis. More recently, their role in several neurodegenerative disorders has been gaining increased momentum as these particles have been shown to promote the spread of toxic factors such as amyloid beta and prions, adding further validity to their role as important regulators of disease pathogenesis. This review briefly summarizes current findings and thoughts on exosome biology in the context of neurodegenerative disorders and the manipulation of these particles for the development of potential therapeutic strategies.

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Qualitative and quantitative analysis of endocytic recycling

Cited by 9 publications

References 18 publications

Quantitative Analysis of Endocytic Recycling of Membrane Proteins by Monoclonal Antibody-Based Recycling Assays

Quantitative Analysis of Endocytic Recycling of Membrane Proteins by Monoclonal Antibody-Based Recycling Assays

Rapid Endosomal Recycling

Emerging roles of extracellular vesicles in neurodegenerative disorders: focus on HIV-associated neurological complications

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