Covalent binding of human thrombin to a human endothelial cell-associated protein

Journal Article · Wed Mar 01 04:00:00 EST 1989 · Experimental Cell Research; (United States)

DOI:https://doi.org/10.1016/0014-4827(89)90177-8· OSTI ID:5623323

Leroy-Viard, K; Jandrot-Perrus, M; Tobelem, G; Guillin, M C ^[1]

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1. Univ. Paris 7 (France)

Binding of {sup 125}I-thrombin to endothelial cells derived from human umbilical vein was studied in tissue culture. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and autoradiography revealed covalent binding of thrombin in a 72-kDa complex. Some characteristics of the 72-kDa complex were compared to those of the complexes formed between thrombin and protease nexin originating from fibroblasts or platelets: (1) its electrophoretic mobility on SDS-PAGE is identical to that of the thrombin-platelet protease nexin complex, (2) heparin prevents the appearance of the complex on the cell surface, (3) plasmin in a 100-fold molar excess prevents the covalent linkage of thrombin, suggesting that the protease specificity of the endothelial component involved in the complex might not be restricted to thrombin. Yet no release, nor any secretion of the endothelial protein, could be detected. These results indicate that active thrombin binds covalently to a specific endothelial protein that is in several respects similar to fibroblasts or platelet protease nexin and provides a thrombin binding site distinct from thrombomodulin and glycosaminoglycans.

• OSTI ID: 5623323
• Journal Information: Experimental Cell Research; (United States), Journal Name: Experimental Cell Research; (United States) Vol. 181:1; ISSN ECREA; ISSN 0014-4827
• Country of Publication: United States
• Language: English

Related Subjects

550301* -- Cytology-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
AMINES
ANIMAL TISSUES
ANIMALS
ANTICOAGULANTS
AUTORADIOGRAPHY
BETA DECAY RADIOISOTOPES
BLOOD COAGULATION FACTORS
BODY
CARBOHYDRATES
CHEMICAL REACTIONS
COAGULANTS
DAYS LIVING RADIOISOTOPES
DRUGS
ELECTRON CAPTURE RADIOISOTOPES
ELECTROPHORESIS
ENDOTHELIUM
ENZYMES
HALOGENATION
HEMATOLOGIC AGENTS
HEMOSTATICS
HEPARIN
HYDROLASES
INTERMEDIATE MASS NUCLEI
INTERNAL CONVERSION RADIOISOTOPES
IODINATION
IODINE 125
IODINE ISOTOPES
ISOTOPES
MAMMALS
MAN
MEMBRANE PROTEINS
MUCOPOLYSACCHARIDES
NUCLEI
ODD-EVEN NUCLEI
ORGANIC COMPOUNDS
ORGANIC SULFUR COMPOUNDS
PEPTIDE HYDROLASES
POLYSACCHARIDES
PRIMATES
PROTEINS
PURIFICATION
RADIOISOTOPES
RECEPTORS
SACCHARIDES
SERINE PROTEINASES
THROMBIN
TISSUES
VERTEBRATES