Please use this identifier to cite or link to this item: http://hdl.handle.net/2445/51226
Title: Ligand binding to heparan sulfate proteoglycans induces their aggregation and distribution along actin cytoskeleton
Author: Marthinho, Rui G.
Castel i Gil, Susanna
Ureña Bares, Jesús Mariano
Fernández Borja, Maria del Mar
Makiya, Ricardo
Olivecrona, Gunilla
Reina del Pozo, Manuel
Alonso, Ana
Vilaró, Senén, 1956-2005
Keywords: Interacció cel·lular
Citosquelet
Proteïnes citosquelètiques
Cell interaction
Cytoskeleton
Cytoskeletal proteins
Issue Date: Nov-1996
Publisher: American Society for Cell Biology
Abstract: Cell surface heparan sulfate proteoglycans (HSPGs) participate in molecular events that regulate cell adhesion, migration, and proliferation. The present study demonstrates that soluble heparin-binding proteins or cross-linking antibodies induce the aggregation of cell surface HSPGs and their distribution along underlying actin filaments. Immunofluorescence and confocal microscopy and immunogold and electron microscopy indicate that, in the absence of ligands, HSPGs are irregularly distributed on the fibroblast cell surface, without any apparent codistribution with the actin cytoskeleton. In the presence of ligand (lipoprotein lipase) or antibodies against heparan sulfate, HSPGs aggregate and colocalize with the actin cytoskeleton. Triton X-100 extraction and immunoelectron microscopy have demonstrated that in this condition HSPGs were clustered and associated with the actin filaments. Crosslinking experiments that use biotinylated lipoprotein lipase have revealed three major proteoglycans as binding sites at the fibroblast cell surface. These cross-linked proteoglycans appeared in the Triton X-100 insoluble fraction. Platinum/carbon replicas of the fibroblast surface incubated either with lipoprotein lipase or antiheparan sulfate showed large aggregates of HSPGs regularly distributed along cytoplasmic fibers. Quantification of the spacing between HSPGs by confocal microscopy confirmed that the nonrandom distribution of HSPG aggregates along the actin cytoskeleton was induced by ligand binding. When cells were incubated either with lipoprotein lipase or antibodies against heparan sulfate, the distance between immunofluorescence spots was uniform. In contrast, the spacing between HSPGs on fixed cells not incubated with ligand was more variable. This highly organized spatial relationship between actin and proteoglycans suggests that cortical actin filaments could organize the molecular machinery involved in signal transduction and molecular movements on the cell surface that are triggered by heparin-binding proteins.
Note: Reproducció del document publicat a: http://dx.doi.org/10.1091/mbc.7.11.1771
It is part of: Molecular Biology of the Cell, 1996, vol. 7, num. 11, p. 1771-1788
Related resource: http://dx.doi.org/10.1091/mbc.7.11.1771
URI: http://hdl.handle.net/2445/51226
ISSN: 1059-1524
Appears in Collections:Articles publicats en revistes (Biologia Cel·lular, Fisiologia i Immunologia)

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