Isolation and Properties of a Thrombin-sensitive Protein of Human Platelets

• Published in: The Journal of biological chemistry 1972-05, Vol.247 (9), p.2723-2731
• Main Authors: Baenziger, Nancy Lewis, Brodie, G.N., Majerus, Philip W.
• Format: Article
• Language: English
• Subjects: Acetates - analysis; Adenosine Triphosphatases - blood; Amino Acids - analysis; Blood Platelets - cytology; Blood Platelets - drug effects; Blood Platelets - enzymology; Carbon Isotopes; Chromatography, Gel; Dansyl Compounds; Electrophoresis; Fluorine; Glucosamine - analysis; Glycoproteins - analysis; Glycoproteins - blood; Glycoproteins - isolation & purification; Humans; Molecular Weight; Neuraminic Acids - analysis; Nitrobenzenes; Sodium Dodecyl Sulfate; Solubility; Subcellular Fractions; Thrombin - pharmacology; Vibration
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1016/S0021-9258(19)45271-X

A glycoprotein of human platelets is released into the incubation medium upon treatment of the intact cells with 1 unit per ml of thrombin for 2 min. The cellular location and properties of this protein differ from those of other thrombin-labile platelet proteins such as fibrin-stabilizing factor, fibrinogen, and the contractile protein thrombosthenin. It occurs only in the particulate fraction and is released only from intact cells. Thrombin concentrations of 100 units per ml do not release this thrombin-sensitive protein from sonicated platelets. As isolated from the incubation supernatant of thrombin-treated platelets the thrombin-sensitive protein is excluded from Sephadex G-200; it has an apparent molecular weight of 190,000 in sodium dodecyl sulfate polyacrylamide gels, from which it has been purified and partially characterized. No smaller subunits have been found. It has limited solubility in the absence of detergent, and is highly sensitive to proteolytic degradation in whole platelets and after release from the cells. Membrane-bound and released forms of the thrombin-sensitive protein show no significant difference in molecular weight or amino acid and carbohydrate composition, suggesting that release does not involve proteolysis of the thrombin-sensitive protein itself by thrombin. This protein contains 3.9% sialic acid, 1.6% N-acetylglucosamine, and 25% each of acidic and nonpolar residues. Although it contains carbohydrate the thrombin-sensitive protein does not represent the major membrane glycoprotein of platelets, for it contains only 5% of the total platelet sialic acid. The thrombin-sensitive protein accounts for approximately 5% of the total protein in the particulate fraction of human platelets and represents about 20 to 30% of the protein found in the incubation supernatant after thrombin treatment.