title: Enhanced receptor-clathrin interactions induced by N-glycan-mediated membrane micropatterning.
abstract: Glycan-protein interactions are emerging as important modulators of membrane protein organization and dynamics, regulating multiple cellular functions. In particular, it has been postulated that glycan-mediated interactions regulate surface residence time of glycoproteins and endocytosis. How this precisely occurs is poorly understood. Here we applied single-molecule-based approaches to directly visualize the impact of glycan-based interactions on the spatiotemporal organization and interaction with clathrin of the glycosylated pathogen recognition receptor dendritic cell-specific intercellular adhesion molecule-3-grabbing nonintegrin (DC-SIGN). We find that cell surface glycan-mediated interactions do not influence the nanoscale lateral organization of DC-SIGN but restrict the mobility of the receptor to distinct micrometer-size membrane regions. Remarkably, these regions are enriched in clathrin, thereby increasing the probability of DC-SIGN-clathrin interactions beyond random encountering. N-glycan removal or neutralization leads to larger membrane exploration and reduced interaction with clathrin, compromising clathrin-dependent internalization of virus-like particles by DC-SIGN. Therefore, our data reveal that cell surface glycan-mediated interactions add another organization layer to the cell membrane at the microscale and establish a novel mechanism of extracellular membrane organization based on the compartments of the membrane that a receptor is able to explore. Our work underscores the important and complex role of surface glycans regulating cell membrane organization and interaction with downstream partners.
authors.affiliation: ICFO-Institut de Ciencies Fotoniques, 08860 Castelldefels, Spain;
authors.forename: Juan A
authors.initials: JA
authors.lastname: Torreno-Pina
authors.affiliation: ICFO-Institut de Ciencies Fotoniques, 08860 Castelldefels, Spain;
authors.forename: Bruno M
authors.initials: BM
authors.lastname: Castro
authors.affiliation: ICFO-Institut de Ciencies Fotoniques, 08860 Castelldefels, Spain;
authors.forename: Carlo
authors.initials: C
authors.lastname: Manzo
authors.affiliation: Department of Tumour Immunology, Radboud Institute for Molecular Life Sciences, Radboud University Medical Center, 6500, HB Nijmegen, The Netherlands;
authors.forename: Sonja I
authors.initials: SI
authors.lastname: Buschow
authors.affiliation: Department of Tumour Immunology, Radboud Institute for Molecular Life Sciences, Radboud University Medical Center, 6500, HB Nijmegen, The Netherlands;Nanobiophysics, MESA+ Institute for Nanotechnology and MIRA Institute for Biomedical Technology and Technical Medicine, University of Twente, 7500 AE, Enschede, The Netherlands; and.
authors.forename: Alessandra
authors.initials: A
authors.lastname: Cambi
authors.affiliation: ICFO-Institut de Ciencies Fotoniques, 08860 Castelldefels, Spain;ICREA-Institució Catalana de Recerca i Estudis Avançats, 08010 Barcelona, Spain maria.garcia-parajo@icfo.es.
authors.forename: Maria F
authors.initials: MF
authors.lastname: Garcia-Parajo
mesh_terms: D000818:Animals; D016466:CHO Cells; D015815:Cell Adhesion Molecules / Q000235:genetics / Q000378:metabolism*; D002966:Clathrin / Q000235:genetics / Q000378:metabolism*; D006224:Cricetinae; D003412:Cricetulus; D006801:Humans; D037181:Lectins, C-Type / Q000235:genetics / Q000378:metabolism*; D021962:Membrane Microdomains / Q000235:genetics / Q000378:metabolism*; D011134:Polysaccharides / Q000378:metabolism*; D011956:Receptors, Cell Surface / Q000235:genetics / Q000378:metabolism*; D000099215:DC-Specific ICAM-3 Grabbing Nonintegrin
chemical_list: D015815:Cell Adhesion Molecules; D002966:Clathrin; D037181:Lectins, C-Type; D011134:Polysaccharides; D011956:Receptors, Cell Surface; D000099215:DC-Specific ICAM-3 Grabbing Nonintegrin
keywords: STED nanoscopy; cell membrane compartmentalization; clathrin-dependent endocytosis; pathogen receptor DC-SIGN; protein glycosylation