Discrimination of deletion to point cytokine mutants based on an array of cross-reactive receptors mimicking protein recognition by heparan sulfate

Abstract Heparan sulfate (HS) is a linear and complex polysaccharide that modulates the biological activities through protein recognition and interaction. Evidence indicates that protein-binding properties of HS are largely dependent on distinctive sulfation and epimerization patterns that are modified by a series of Golgi-localized enzymes. In particular, the glucuronyl C5-epimerase (Hsepi) converts D-glucuronic acid (GlcA) residues to L-iduronic acid (IdoA) and 2-O-sulfotransferase (2OST) catalyzes sulfation at C2 position of IdoA and rarely GlcA residues. Mice lacking both Hsepi and 2OST display multiple development defects, indicating the importance of IdoA in HS. Here, to gain greater insights of HS structure–function relationships, as well as a better understanding of the regulatory mechanisms of Hsepi and 2OST, the fine structure and cellular signaling functions of HS were investigated after restoration of Hsepi in the mutant mouse embryonic fibroblast (MEF) cells. Introduction of Hsepi into the Hsepi mutant MEF cells led to robustly increased proportion of IdoA residues, which rescued the cell signaling in responding to fibroblast growth factor 2. However, we found that Hsepi knockout had no influence on either cellular transportation or enzymatic activity of 2OST in the MEF cells, which is not in accord with the findings reported, suggesting that the enzymatic activity and cellular transportation of 2OST and Hsepi might be differently regulated.

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Localization of cell-surface and basement-membrane heparan-sulfate proteoglycans using the malaria circumsporozoite protein

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100169201 ◽

1994 ◽

Vol 52 ◽

pp. 294-295

Author(s):

U. Frevert ◽

S. Sinnis ◽

C. Cerami ◽

V. Nussenzweig

Keyword(s):

Heparan Sulfate ◽

Protein A ◽

Kidney Tissue ◽

Plasmodium Species ◽

Its Region ◽

Basement Membranes ◽

Heparan Sulfate Proteoglycans ◽

Infected Mosquito ◽

Complement Components ◽

Region Ii

Malaria sporozoites, which invade hepatocytes within minutes after transmission by an infected mosquito, are covered with the circumsporozoite (CS) protein, which in all Plasmodium species contains the conserved region II-plus. This region is also found as a cell-adhesive motif in a variety of host proteins like thrombospondin, properdin and the terminal complement components.The CS protein with its region II-plus specifically binds to heparan sulfate proteoglycans (HSPG) on the basolateral surface of hepatocytes in the space of Disse (FIG. 1), to certain basolateral cell membranes and basement membranes of the kidney (FIG. 2) as well as to heparin in the granules of connective tissue mast cells. The distribution of the HSPG receptors for the CS protein was examined by incubation of Lowicryl K4M or LR White sections of liver and kidney tissue with the recombinant CS ligand, whose binding sites were detected with a monoclonal anti-CS antibody and protein A gold.

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3-O-sulfation of heparan sulfate modulates breast cancer cell invasiveness by enhancing signaling via the map kinase- and TCF4 (TCF7L2) pathways

Experimental and Clinical Endocrinology & Diabetes ◽

10.1055/s-0035-1547653 ◽

2015 ◽

Vol 122 (03) ◽

Author(s):

AV Kumar ◽

ES Gassar ◽

L Kiesel ◽

G Yip ◽

M Götte

Keyword(s):

Breast Cancer ◽

Heparan Sulfate ◽

Map Kinase ◽

Cancer Cell ◽

Breast Cancer Cell ◽

Cell Invasiveness ◽

Cancer Cell Invasiveness

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The heparan sulfate sulfotransferase HS2ST1 modulates breast cancer cell invasiveness via the EGFR and MAPK signaling pathways

Experimental and Clinical Endocrinology & Diabetes ◽

10.1055/s-0035-1547652 ◽

2015 ◽

Vol 122 (03) ◽

Author(s):

AV Kumar ◽

S Katakam ◽

L Kiesel ◽

M Götte

Keyword(s):

Breast Cancer ◽

Heparan Sulfate ◽

Signaling Pathways ◽

Cancer Cell ◽

Breast Cancer Cell ◽

Mapk Signaling ◽

Cell Invasiveness ◽

Cancer Cell Invasiveness ◽

Mapk Signaling Pathways

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Specific Cross-reaction of IgG Anti-phospholipid Antibody with Platelet Glycoprotein IIIa

Thrombosis and Haemostasis ◽

10.1055/s-0038-1650238 ◽

1996 ◽

Vol 75 (01) ◽

pp. 168-174 ◽

Cited By ~ 13

Author(s):

Shigeru Tokita ◽

Morio Arai ◽

Naomasa Yamamoto ◽

Yasuhiro Katagiri ◽

Kenjiro Tanoue ◽

...

Keyword(s):

Platelet Aggregation ◽

Heparan Sulfate ◽

Disulfide Bonds ◽

Dextran Sulfate ◽

Human Platelets ◽

Platelet Glycoprotein ◽

Activated Platelets ◽

Fibrinogen Binding ◽

Dose Dependent Fashion ◽

Glycoprotein Iiia

SummaryTo study the pathological functions of anti-phospholipid (anti-PL) antibodies, we have analyzed their effect on platelet function. We identified an IgG anti-PL mAb, designated PSG3, which cross-reacted specifically with glycoprotein (GP) IIIa in human platelets and inhibited platelet aggregation. PSG3 bound also to certain polyanionic substances, such as double-stranded DNA, heparan sulfate, dextran sulfate and acetylated-LDL, but not to other polyanionic substances. The binding of PSG3 to GPIIIa was completely inhibited by heparan sulfate and dextran sulfate, indicating that PSG3 recognizes a particular array of negative charges expressed on both GPIIIa and the specified polyanionic substances. Since neither neuraminidase- nor endoglycopeptidase F-treatment of GPIIIa had any significant effect on the binding of PSG3, this array must be located within the amino acid sequence of GPIIIa but not in the carbohydrate moiety. Reduction of the disulfide bonds in GPIIIa greatly reduced its reactivity, suggesting that the negative charges in the epitope are arranged in a particular conformation. PSG3 inhibited platelet aggregation induced by either ADP or collagen, it also inhibited fibrinogen binding to activated platelets in a dose-dependent fashion. PSG3, however, did not inhibit the binding of GRGDSP peptide to activated platelets. These results suggest that the PSG3 epitope on GPIIIa contains a particular array of negative charges, and possibly affects the fibrinogen binding to GPIIb/IIIa complex necessary for platelet aggregation.

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