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Evaluating the Role of Galectins in Clathrin-Independent Endocytosis

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Galectins

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2442))

Abstract

Galectin-3 is a chimeric galectin involved in diverse intracellular and extracellular functions. Galectin-3 is synthesized in the cytoplasm and then released extracellularly by a poorly understood non-canonical secretion mechanism. As a result, it can play important roles both inside and outside the cell. One important extracellular role of galectin-3 is in modulating clathrin-independent endocytosis (CIE), a form of cellular internalization that is still not well understood. CIE, unlike clathrin-mediated endocytosis, has neither defined signaling sequences nor cytoplasmic machinery. As a result, extracellular interactions like the galectin-glycan interactions are thought to directly drive changes in CIE. This chapter discusses the methods designed to study the role of galectin-glycan interactions in CIE, which have provided us with insight into the functions of galectin-3 and cell surface glycans during CIE cargo internalization. These methods include media supplementation for metabolic glycoengineering, antibody internalization assays, lectin panels to assay changes in glycan patterns, exogenous galectin-3 supplementation, galectin-3 secretion assays, and in vitro assays to monitor the effect of galectins on CIE.

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Author information

Authors and Affiliations

  1. National Institute of Diabetes and Digestive and Kidney Diseases, Bethesda, MD, USA

    Mohit P. Mathew & John A. Hanover

  2. National Heart, Lung and Blood Institute, Bethesda, MD, USA

    Julie G. Donaldson

Authors
  1. Mohit P. Mathew
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  2. Julie G. Donaldson
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  3. John A. Hanover
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Corresponding author

Correspondence to Mohit P. Mathew .

Editor information

Editors and Affiliations

  1. Joint Program in Transfusion Medicine Brigham and Women’s Hospital, Harvard Medical School, Boston, MA, USA

    Sean R. Stowell

  2. Joint Program in Transfusion Medicine Brigham and Women’s Hospital, Harvard Medical School, Boston, MA, USA

    Connie M. Arthur

  3. Department of Surgery Beth Israel Deaconess Medical Center National Center for Functional Glycomics, Harvard Medical School, Boston, MA, USA

    Richard D. Cummings

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Mathew, M.P., Donaldson, J.G., Hanover, J.A. (2022). Evaluating the Role of Galectins in Clathrin-Independent Endocytosis. In: Stowell, S.R., Arthur, C.M., Cummings, R.D. (eds) Galectins. Methods in Molecular Biology, vol 2442. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2055-7_21

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  • DOI: https://doi.org/10.1007/978-1-0716-2055-7_21

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2054-0

  • Online ISBN: 978-1-0716-2055-7

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