Modulation of glycosaminoglycan production and antithrombin III binding by cultured aortic endothelial cells treated with 4-methylumbelliferyl-beta-D-xyloside
The interaction between antithrombin III and heparinlike glycosaminoglycan molecules present on the vascular surface seems to play a significant role in the regulation of coagulation. We have tested the hypothesis that altered synthesis of glycosaminoglycans by endothelial cells could influence this interaction by using 4-methylumbelliferyl-beta-D-xyloside for metabolic perturbation of glycosaminoglycan production. Incubation of purified porcine /sup 125/I-antithrombin III with cultured porcine aortic endothelial cells demonstrated specific, time-dependent, saturable binding of this protease inhibitor to the endothelial cell surface with antithrombin III concentration at half-maximal binding of approximately 40 nM. This binding was displaced by heparin and was completely abolished by selective removal of heparan sulfate from cells with heparitinase, indicating that antithrombin III binds to heparan sulfate on the surface of endothelial cells. Incubation of cell cultures with beta-D-xyloside resulted in a reduction of maximum antithrombin III binding by approximately 65% with little alteration in binding affinity. beta-D-Xyloside did not affect the cellular growth or morphology. Reduction of the binding after exposure to various concentrations (33 to 500 microM) of xyloside occurred in parallel with the decrease in incorporation of both /sup 35/S-sulfate and /sup 3/H-glucosamine into cell surface heparan sulfate. Whereas the size of heparan sulfate chains derived from the cell surface was not altered by xyloside treatment, they appeared to have slightly less net negative charge and a significantly reduced proportion of the molecule with high affinity for antithrombin III in the presence of xyloside.
- Research Organization:
- Kochi Medical School, Japan
- OSTI ID:
- 5607684
- Journal Information:
- Arteriosclerosis (Dallas); (United States), Journal Name: Arteriosclerosis (Dallas); (United States) Vol. 7:6; ISSN ARTRD
- Country of Publication:
- United States
- Language:
- English
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59 BASIC BIOLOGICAL SCIENCES
AMINES
ANIMAL TISSUES
ANIMALS
ANTICOAGULANTS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIOCHEMICAL REACTION KINETICS
BIOSYNTHESIS
BODY
CARBOHYDRATES
CELL CONSTITUENTS
CELL MEMBRANES
COAGULANTS
DAYS LIVING RADIOISOTOPES
DOMESTIC ANIMALS
DRUGS
ELECTRON CAPTURE RADIOISOTOPES
ENDOTHELIUM
ENZYME INHIBITORS
ENZYMES
EVEN-ODD NUCLEI
GLUCOPROTEINS
GLUCOSAMINE
HEMATOLOGIC AGENTS
HEMOSTATICS
HEPARIN
HEXOSAMINES
HEXOSES
HYDROLASES
INTERMEDIATE MASS NUCLEI
IODINE 125
IODINE ISOTOPES
ISOTOPE APPLICATIONS
ISOTOPES
KINETICS
LABELLED COMPOUNDS
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MAMMALS
MEMBRANES
MONOSACCHARIDES
MUCOPOLYSACCHARIDES
NUCLEI
ODD-EVEN NUCLEI
ORGANIC COMPOUNDS
ORGANIC SULFUR COMPOUNDS
OXYGEN COMPOUNDS
PEPTIDE HYDROLASES
POLYSACCHARIDES
PROTEINS
RADIOISOTOPES
REACTION KINETICS
RESPONSE MODIFYING FACTORS
SACCHARIDES
SERINE PROTEINASES
SULFATES
SULFUR 35
SULFUR COMPOUNDS
SULFUR ISOTOPES
SWINE
SYNTHESIS
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TIME DEPENDENCE
TISSUES
TRACER TECHNIQUES
TRITIUM COMPOUNDS
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