Heparanase and a Synthetic Peptide of Heparan Sulfate-interacting Protein Recognize Common Sites on Cell Surface and Extracellular Matrix Heparan Sulfate

Cell surface heparan sulfate proteoglycan (HSPG) from metastatic mouse melanoma cells initiates cell adhesion to the synthetic peptide FN-C/H II, a heparin-binding peptide from the 33-kD A chain-derived fragment of fibronectin. Mouse melanoma cell adhesion to FN-C/H II was sensitive to soluble heparin and pretreatment of mouse melanoma cells with heparitinase. In contrast, cell adhesion to the fibronectin synthetic peptide CS1 is mediated through an alpha 4 beta 1 integrin and was resistant to heparin or heparitinase treatment. Mouse melanoma cell HSPG was metabolically labeled with [35S]sulfate and extracted with detergent. After HPLC-DEAE purification, 35S-HSPG eluted from a dissociative CL-4B column with a Kav approximately 0.45, while 35S-heparan sulfate (HS) chains eluted with a Kav approximately 0.62. The HSPG contained a major 63-kD core protein after heparitinase digestion. Polyclonal antibodies generated against HSPG purified from mouse melanoma cells grown in vivo also identified a 63-kD core protein. This HSPG is an integral plasma membrane component by virtue of its binding to Octyl Sepharose affinity columns and that anti-HSPG antibody staining exhibited a cell surface localization. The HSPG is anchored to the cell surface through phosphatidylinositol (PI) linkages, as evidenced in part by the ability of PI-specific phospholipase C to eliminate binding of the detergent-extracted HSPG to Octyl Sepharose. Furthermore, the mouse melanoma HSPG core protein could be metabolically labeled with 3H-ethanolamine. The involvement of mouse melanoma cell surface HSPG in cell adhesion to fibronectin was also demonstrated by the ability of anti-HSPG antibodies and anti-HSPG IgG Fab monomers to inhibit mouse melanoma cell adhesion to FN-C/H II. 35S-HSPG and 35S-HS bind to FN-C/H II affinity columns and require 0.25 M NaCl for elution. However, heparitinase-treated 125I-labeled HSPG failed to bind FN-C/H II, suggesting that HS, and not HSPG core protein, binds FN-C/H II. These data support the hypothesis that a phosphatidylinositol-anchored HSPG on mouse melanoma cells (MPIHP-63) initiates recognition to FN-C/H II, and implicate PI-associated signal transduction pathways in mediating melanoma cell adhesion to this defined ligand.

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Targeting heparin and heparan sulfate protein interactions

Organic & Biomolecular Chemistry ◽

10.1039/c7ob01058c ◽

2017 ◽

Vol 15 (27) ◽

pp. 5656-5668 ◽

Cited By ~ 44

Author(s):

Ryan J. Weiss ◽

Jeffrey D. Esko ◽

Yitzhak Tor

Keyword(s):

Extracellular Matrix ◽

Cell Surface ◽

Heparan Sulfate ◽

Protein Interactions ◽

Human Disease ◽

Binding Proteins ◽

Cellular Functions ◽

Animal Cells ◽

Carbohydrate Chains

Heparan sulfate is ubiquitously expressed on the cell surface and in the extracellular matrix of all animal cells. These negatively-charged carbohydrate chains play essential roles in many important cellular functions by interacting with various heparan sulfate binding proteins (HSBP). This review discusses methods for targeting these complex biomolecules, as strategies for treating human disease.

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Alveolar type II cells synthesize hydrophobic cell-associated proteoglycans with multiple core proteins

AJP Lung Cellular and Molecular Physiology ◽

10.1152/ajplung.1992.263.3.l348 ◽

1992 ◽

Vol 263 (3) ◽

pp. L348-L356 ◽

Cited By ~ 2

Author(s):

W. M. Maniscalco ◽

M. H. Campbell

Keyword(s):

Extracellular Matrix ◽

Cell Surface ◽

Heparan Sulfate ◽

Core Protein ◽

Primary Cultures ◽

Alveolar Type ◽

Type Ii Cells ◽

Type Ii ◽

Alveolar Type Ii Cells ◽

Core Proteins

Type II alveolar epithelial cells interact with the extracellular matrix via cell surface receptors for matrix ligands. Cell surface proteoglycans, which are hydrophobic due to their membrane insertion domains, are one of several classes of molecules that may be receptors for matrix ligands. To analyze the hydrophobic proteoglycans synthesized by adult alveolar type II cells, we labeled these cells with 35SO4 and [3H]leucine in short-term primary cultures. Cell-associated hydrophobic proteoglycans and culture medium-derived proteoglycans were purified and characterized. Both the hydrophobic proteoglycans and medium-derived proteoglycans, which were not hydrophobic, had mainly heparan sulfate glycosaminoglycans. Analysis of core proteins of the hydrophobic proteoglycans showed three proteins, 47, 65, and 90 kDa. The 47- and 65-kDa core proteins were substituted only with heparan sulfate chains. The 90-kDa core protein was seen only after digestion with both heparitinase and chondroitin ABC lyase, suggesting it was a hybrid having both heparan sulfate and chondroitin-dermatan sulfate chains. These findings were confirmed by iodination of the core proteins. The hydrophobic cell-associated proteoglycans inserted into artificial liposomes, whereas the medium-derived molecules did not. These data document heterogeneity in core protein and glycosaminoglycan chains among hydrophobic proteoglycans synthesized in vitro by adult alveolar type II cells. These molecules may have diverse functions in regulating type II cell interaction with the extracellular matrix.

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Cell Surface Localization of Heparanase on Macrophages Regulates Degradation of Extracellular Matrix Heparan Sulfate

The Journal of Immunology ◽

10.4049/jimmunol.172.6.3830 ◽

2004 ◽

Vol 172 (6) ◽

pp. 3830-3835 ◽

Cited By ~ 55

Author(s):

Norihiko Sasaki ◽

Nobuaki Higashi ◽

Tomohiro Taka ◽

Motowo Nakajima ◽

Tatsuro Irimura

Keyword(s):

Extracellular Matrix ◽

Cell Surface ◽

Heparan Sulfate ◽

Surface Localization ◽

Cell Surface Localization

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The heparan sulfate and its diverse biological activities

Current Issues in Pharmacy and Medical Sciences ◽

10.1515/cipms-2015-0016 ◽

2014 ◽

Vol 27 (4) ◽

pp. 209-212

Author(s):

Iwona Kaznowska-Bystryk

Keyword(s):

Extracellular Matrix ◽

Growth Factors ◽

Cell Surface ◽

Heparan Sulfate ◽

Biological Function ◽

Molecular Level ◽

Biological Activities ◽

Therapeutic Strategies ◽

Detailed Knowledge ◽

And Storage

Abstract Heparan sulfate (HS) is one of the most common glycosaminoglycan (GAG) in mammals. It is composed of relatively simple disaccharide subunits, which, by further modification, such as sulfation and epimerization, potentially offer huge diversity in biological function. GAG chains of different length, different patterns of sulfation, and other modifications, depending on location, generate unique forms. Due to polyanion charges, these compounds can interact with other molecules, such as proteins, cytokines, chemokines and growth factors, both on the cell surface and inside the extracellular matrix. These interactions serve protective and storage functions for the compounds, safeguarding them from proteolysis. In this way, HS is involved in numerous signaling pathways, and in growth and differentiation processes. Disrupted interactions between the HS and growth factors, cytokines or other proteins have been observed in various disorders, among these Alzheimer’s disease, epilepsy, atherosclerosis, diabetes, and cancer processes. Detailed knowledge of these relationships at the molecular level will allow researchers to understand the mechanisms underlying these disorders and enable the development of effective therapeutic strategies.

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Heparan Sulfate Proteoglycans and Viral Attachment: True Receptors or Adaptation Bias?

Viruses ◽

10.3390/v11070596 ◽

2019 ◽

Vol 11 (7) ◽

pp. 596 ◽

Cited By ~ 59

Author(s):

Cagno ◽

Tseligka ◽

Jones ◽

Tapparel

Keyword(s):

Extracellular Matrix ◽

Cell Surface ◽

Heparan Sulfate ◽

Blood Coagulation ◽

Biological Activities ◽

Extracellular Matrix Proteins ◽

Heparan Sulfate Proteoglycans ◽

Matrix Proteins ◽

Cell Homeostasis ◽

Viral Attachment

Heparan sulfate proteoglycans (HSPG) are composed of unbranched, negatively charged heparan sulfate (HS) polysaccharides attached to a variety of cell surface or extracellular matrix proteins. Widely expressed, they mediate many biological activities, including angiogenesis, blood coagulation, developmental processes, and cell homeostasis. HSPG are highly sulfated and broadly used by a range of pathogens, especially viruses, to attach to the cell surface.

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Relationship of heparan sulfate proteoglycans to the cytoskeleton and extracellular matrix of cultured fibroblasts.

The Journal of Cell Biology ◽

10.1083/jcb.99.5.1743 ◽

1984 ◽

Vol 99 (5) ◽

pp. 1743-1753 ◽

Cited By ~ 106

Author(s):

A Woods ◽

M Höök ◽

L Kjellén ◽

C G Smith ◽

D A Rees

Keyword(s):

Extracellular Matrix ◽

Cell Surface ◽

Heparan Sulfate ◽

Focal Adhesions ◽

Dermal Fibroblasts ◽

Heparan Sulfate Proteoglycans ◽

Immunofluorescent Staining ◽

Rat Embryo ◽

Double Labeling ◽

Cultured Fibroblasts

The distribution of heparan sulfate proteoglycans (HSPG) on cultured fibroblasts was monitored using an antiserum raised against cell surface HSPG from rat liver. After seeding, HSPG was detected by immunofluorescence first on cell surfaces and later in fibrillar deposits of an extracellular matrix. Cell surface HSPG aligned with microfilament bundles of rat embryo fibroblasts seen by phase-contrast microscopy but was diffuse on transformed rat dermal fibroblasts (16C cells) which lack obvious stress fibers. Focal adhesions isolated from either cell type and monitored by interference reflection microscopy showed a concentration of HSPG labeling with respect to the rest of the membrane. Increased labeling in these areas was also seen for fibronectin (FN) by using an antiserum that detects both plasma and cell-derived FN. Double immunofluorescent staining of fully adherent rat embryo fibroblast cells showed some co-distribution of HSPG and FN, and this was confirmed by immunoelectron microscopy, which detected HSPG at localized areas of dorsal and ventral cell membranes, overlapping cell margins, and in the extracellular matrix. During cell shape changes on rounding and spreading, HSPG and FN may not co-distribute. Double labeling for actin and either HSPG or FN showed a closer correlation of actin with HSPG than with FN. The studies are consistent with HSPG being closely involved in a transmembrane cytoskeletal-matrix interaction; the possibility that HSPG coordinates the deposition of FN and other matrix components with cytoskeletal organization is discussed.

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Regulation of Notch signaling by Drosophila heparan sulfate 3-O sulfotransferase

The Journal of Cell Biology ◽

10.1083/jcb.200403077 ◽

2004 ◽

Vol 166 (7) ◽

pp. 1069-1079 ◽

Cited By ~ 57

Author(s):

Keisuke Kamimura ◽

John M. Rhodes ◽

Ryu Ueda ◽

Melissa McNeely ◽

Deepak Shukla ◽

...

Keyword(s):

Extracellular Matrix ◽

Drosophila Melanogaster ◽

Cell Surface ◽

Heparan Sulfate ◽

Notch Signaling ◽

Intracellular Trafficking ◽

Specific Binding ◽

Notch Pathway ◽

Notch Protein ◽

Sulfation Pattern

Heparan sulfate (HS) regulates the activity of various ligands and is involved in molecular recognition events on the cell surface and in the extracellular matrix. Specific binding of HS to different ligand proteins depends on the sulfation pattern of HS. For example, the interaction between antithrombin and a particular 3-O sulfated HS motif is thought to modulate blood coagulation. However, a recent study of mice defective for this modification suggested that 3-O sulfation plays other biological roles. Here, we show that Drosophila melanogaster HS 3-O sulfotransferase-b (Hs3st-B), which catalyzes HS 3-O sulfation, is a novel component of the Notch pathway. Reduction of Hs3st-B function by transgenic RNA interference compromised Notch signaling, producing neurogenic phenotypes. We also show that levels of Notch protein on the cell surface were markedly decreased by loss of Hs3st-B. These findings suggest that Hs3st-B is involved in Notch signaling by affecting stability or intracellular trafficking of Notch protein.

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