Heparan Sulfate Proteoglycans Are Receptors for the Cell-surface Trafficking and Biological Activity of Transglutaminase-2

AbstractTranscellular propagation of aggregate “seeds” has been proposed to mediate progression of neurodegenerative diseases in tauopathies and α-synucleinopathies. We have previously determined that tau and α-synuclein aggregates bind heparan sulfate proteoglycans (HSPGs) on the cell surface. This mediates uptake and intracellular seeding. The specificity and mode of binding to HSPGs has been unknown. We used modified heparins to determine the size and sulfation requirements of glycosaminoglycan (GAGs) binding to aggregates in biochemical and cell uptake and seeding assays. Aggregates of tau require a precise GAG architecture with defined sulfate moieties in the N- and 6-O-positions, whereas α-synuclein and Aβ rely slightly more on overall charge on the GAGs. To determine the genetic requirements for aggregate uptake, we individually knocked out the major genes of the HSPG synthesis pathway using CRISPR/Cas9 in HEK293T cells. Knockout of EXT1, EXT2 and EXTL3, N-sulfotransferase (NDST1), and 6-O-sulfotransferase (HS6ST2) significantly reduced tau uptake. α-Synuclein was not sensitive to HS6ST2 knockout. Good correlation between pharmacologic and genetic manipulation of GAG binding by tau and α-synuclein indicates specificity that may help elucidate a path to mechanism-based inhibition of transcellular propagation of pathology.

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Cooperation of Syndecan-2 and Syndecan-4 among Cell Surface Heparan Sulfate Proteoglycans in the Actin Cytoskeletal Organization of Lewis Lung Carcinoma Cells

The Journal of Biochemistry ◽

10.1093/jb/mvh015 ◽

2004 ◽

Vol 135 (1) ◽

pp. 129-137 ◽

Cited By ~ 27

Author(s):

Y. Kusano

Keyword(s):

Cell Surface ◽

Heparan Sulfate ◽

Lung Carcinoma ◽

Lewis Lung Carcinoma ◽

Heparan Sulfate Proteoglycans ◽

Carcinoma Cells ◽

Lewis Lung ◽

Cytoskeletal Organization ◽

Lung Carcinoma Cells ◽

Lewis Lung Carcinoma Cells

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Heparan Sulfate Proteoglycans May Promote or Inhibit Cancer Progression by Interacting with Integrins and Affecting Cell Migration

BioMed Research International ◽

10.1155/2015/453801 ◽

2015 ◽

Vol 2015 ◽

pp. 1-8 ◽

Cited By ~ 24

Author(s):

Mariana A. Soares ◽

Felipe C. O. B. Teixeira ◽

Miguel Fontes ◽

Ana Lúcia Arêas ◽

Marcelo G. Leal ◽

...

Keyword(s):

Cell Adhesion ◽

Cell Surface ◽

Heparan Sulfate ◽

Cancer Progression ◽

Heparan Sulfate Proteoglycans ◽

Transmembrane Receptors ◽

Surface Molecules ◽

Cell Matrix ◽

Cell Organization ◽

The Relationship

The metastatic disease is one of the main consequences of tumor progression, being responsible for most cancer-related deaths worldwide. This review intends to present and discuss data on the relationship between integrins and heparan sulfate proteoglycans in health and cancer progression. Integrins are a family of cell surface transmembrane receptors, responsible for cell-matrix and cell-cell adhesion. Integrins’ main functions include cell adhesion, migration, and survival. Heparan sulfate proteoglycans (HSPGs) are cell surface molecules that play important roles as cell receptors, cofactors, and overall direct or indirect contributors to cell organization. Both molecules can act in conjunction to modulate cell behavior and affect malignancy. In this review, we will discuss the different contexts in which various integrins, such asα5,αV,β1, andβ3, interact with HSPGs species, such as syndecans and perlecans, affecting tissue homeostasis.

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BETA2/NeuroD Protein Transduction Requires Cell Surface Heparan Sulfate Proteoglycans

Human Gene Therapy ◽

10.1089/hum.2007.18.ft-268 ◽

2006 ◽

Vol 0 (0) ◽

pp. 061206073830001

Author(s):

Hirofumi Noguchi ◽

Michiko Ueda ◽

Shinichi Matsumoto ◽

Naoya Kobayashi ◽

Shuji Hayashi

Keyword(s):

Cell Surface ◽

Heparan Sulfate ◽

Heparan Sulfate Proteoglycans ◽

Protein Transduction

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Interaction of an Outer Membrane Protein of Enterotoxigenic Escherichia coli with Cell Surface Heparan Sulfate Proteoglycans

Infection and Immunity ◽

10.1128/iai.70.3.1530-1537.2002 ◽

2002 ◽

Vol 70 (3) ◽

pp. 1530-1537 ◽

Cited By ~ 44

Author(s):

James M. Fleckenstein ◽

James T. Holland ◽

David L. Hasty

Keyword(s):

Escherichia Coli ◽

Epithelial Cell ◽

Epithelial Cells ◽

Cell Surface ◽

Heparan Sulfate ◽

Heparan Sulfate Proteoglycans ◽

Eukaryotic Cells ◽

Heparin Binding ◽

Wild Type ◽

E Coli

ABSTRACT We have previously shown that enterotoxigenic invasion protein A (Tia), a 25-kDa outer membrane protein encoded on an apparent pathogenicity island of enterotoxigenic Escherichia coli (ETEC) strain H10407, mediates attachment to and invasion into cultured human gastrointestinal epithelial cells. The epithelial cell receptor(s) for Tia has not been identified. Here we show that Tia interacts with cell surface heparan sulfate proteoglycans. Recombinant E. coli expressing Tia mediated invasion into wild-type epithelial cell lines but not invasion into proteoglycan-deficient cells. Furthermore, wild-type eukaryotic cells, but not proteoglycan-deficient eukaryotic cells, attached to immobilized polyhistidine-tagged recombinant Tia (rTia). Binding of epithelial cells to immobilized rTia was inhibited by exogenous heparan sulfate glycosaminoglycans but not by hyaluronic acid, dermatan sulfate, or chondroitin sulfate. Similarly, pretreatment of eukaryotic cells with heparinase I, but not pretreatment of eukaryotic cells with chrondroitinase ABC, inhibited attachment to rTia. In addition, we also observed heparin binding to both immobilized rTia and recombinant E. coli expressing Tia. Heparin binding was inhibited by a synthetic peptide representing a surface loop of Tia, as well as by antibodies directed against this peptide. Additional studies indicated that Tia, as a prokaryotic heparin binding protein, may also interact via sulfated proteoglycan molecular bridges with a number of mammalian heparan sulfate binding proteins. These findings suggest that the binding of Tia to host epithelial cells is mediated at least in part through heparan sulfate proteoglycans and that ETEC belongs on the growing list of pathogens that utilize these ubiquitous cell surface molecules as receptors.

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Cell Surface Heparan Sulfate Proteoglycans Mediate the Internalization of PDX-1 Protein

Cell Transplantation ◽

10.3727/000000008783906892 ◽

2008 ◽

Vol 17 (1-2) ◽

pp. 91-97 ◽

Cited By ~ 10

Author(s):

Michiko Ueda ◽

Shinichi Matsumoto ◽

Shuji Hayashi ◽

Naoya Kobayashi ◽

Hirofumi Noguchi

Keyword(s):

Cell Surface ◽

Heparan Sulfate ◽

Heparan Sulfate Proteoglycans

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Dual interaction of the malaria circumsporozoite protein with the low density lipoprotein receptor-related protein (LRP) and heparan sulfate proteoglycans.

Journal of Experimental Medicine ◽

10.1084/jem.184.5.1699 ◽

1996 ◽

Vol 184 (5) ◽

pp. 1699-1711 ◽

Cited By ~ 58

Author(s):

M Shakibaei ◽

U Frevert

Keyword(s):

Cell Surface ◽

Heparan Sulfate ◽

Low Density Lipoprotein ◽

Density Lipoprotein ◽

Heparan Sulfate Proteoglycans ◽

Low Density ◽

Lipoprotein Receptor ◽

Related Protein ◽

Density Lipoprotein Receptor ◽

Lipoprotein Receptor Related Protein

Speed and selectivity of hepatocyte invasion by malaria sporozoites have suggested a receptor-mediated mechanism and the specific interaction of the circumsporozoite (CS) protein with liver-specific heparan sulfate proteoglycans (HSPGs) has been implicated in the targeting to the liver. Here we show that the CS protein interacts not only with cell surface heparan sulfate, but also with the low density lipoprotein receptor-related protein (LRP). Binding of 125I-CS protein to purified LRP occurs with a Kd of 4.9 nM and can be inhibited by the receptor-associated protein (RAP). Blockage of LRP by RAP or anti-LRP antibodies on heparan sulfate-deficient CHO cells results in more than 90% inhibition of binding and endocytosis of recombinant CS protein. Conversely, blockage or enzymatic removal of the cell surface heparan sulfate from LRP-deficient embryonic mouse fibroblasts yields the same degree of inhibition. Heparinase-pretreatment of LRP-deficient fibroblasts or blockage of LRP on heparan sulfate-deficient CHO cells by RAP, lactoferrin, or anti-LRP antibodies reduces Plasmodium berghei invasion by 60-70%. Parasite development in heparinase-pretreated HepG2 cells is inhibited by 65% when RAP is present during sporozoite invasion. These findings suggest that malaria sporozoites utilize the interaction of the CS protein with HSPGs and LRP as the major mechanism for host cell invasion.

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