Assay of Fibrinopeptides A and B and of Released Platelet Proteins as a Measure of Activation of Hemostasis

Summary1. Platelet fibrinogen has been purified from washed bovine platelets. The procedure was based on the methods for purification of plasma fibrinogen by fractionated precipitations and extractions with ethanol and glycine below 0°, and precipitation of proteins by dimethylformamide at 0°.2. The platelet extract obtained by freezing and thawing of the cells, freed from insoluble material by centrifugation at 23,000 x g for 30 min, contained 0.22 ±0.003mg fibrinogen per 109 platelets. Total protein of this fraction was 0.77 ±0.08 mg per 109 platelets whereas that of the insoluble fraction was 0.79 ±0.09 mg per 109 platelets.3. The most purified platelet fibrinogen fraction contained 91-98% of the protein in a thrombin-clottable state. The yield was approx. 20%. It showed homogeneity in analytical ultracentrifugation, in immunoelectrophoresis using an antiserum produced by immunization of rabbits against platelet extract, and in starch gel electrophoresis using a discontinuous system of Tris HCl and borate buffers offering a high resolution power towards the platelet proteins. Polyacrylamide disc electrophoresis revealed two to three faint lines behind the main fibrinogen line. At least one such line was also observed with purified plasma fibrinogen.

Download Full-text

Bovine Platelet Proteins

Thrombosis and Haemostasis ◽

10.1055/s-0038-1653555 ◽

1969 ◽

Vol 21 (03) ◽

pp. 409-418 ◽

Cited By ~ 2

Author(s):

S Łopaciuk ◽

N. O Solum

Keyword(s):

Bovine Serum ◽

Protein Composition ◽

Disc Electrophoresis ◽

Plasma Fibrinogen ◽

Plasma Albumin ◽

Precipitin Line ◽

Bovine Plasma ◽

Platelet Proteins

Summary1. The protein composition of bovine platelet extracts has been investigated by immunoelectrophoresis and polyacrylamide disc electrophoresis. The information obtained is discussed as a basis for study on platelet fibrinogen.2. With antiserum to platelet proteins 11 precipitin lines were observed 3 of which corresponded electrophoretically to plasma albumin, fibrinogen and γ-globulin. These lines were not seen using the same antiserum absorbed with bovine plasma. The 8 additional lines were still present indicating that they represented specific platelet components. Antiserum to plasma produced the 3 above-mentioned lines, but no others.3. With antiserum to purified bovine plasma fibrinogen 3 precipitin lines were observed. The fibrinogen line was the dominant one. The 2 additional lines did not disappear by absorption of the antiserum with bovine serum nor by incubation of the extracts with thrombin. The latter treatment totally removed the fibrinogen line.4. A non-fibrinogen precipitin line, observed only with the antiserum to platelet extract and positioned in the β2-globulin region, disappeared by the incubation of platelet extracts with thrombin.

Download Full-text

Diversity of Glycoprotein Deficiencies in Glanzmann’s Thrombasthenia

Thrombosis and Haemostasis ◽

10.1055/s-0038-1660085 ◽

1985 ◽

Vol 54 (03) ◽

pp. 626-629 ◽

Cited By ~ 4

Author(s):

M Meyer ◽

F H Herrmann

Keyword(s):

High Resolution ◽

Gel Electrophoresis ◽

Type Ii ◽

Two Dimensional ◽

Two Dimensional Gel Electrophoresis ◽

Structural Variant ◽

Glanzmann’S Thrombasthenia ◽

Crossed Immunoelectrophoresis ◽

Glanzmann's Thrombasthenia ◽

Platelet Proteins

SummaryThe platelet proteins of 9 thrombasthenic patients from 7 families were analysed by high resolution two-dimensional gel electrophoresis (HR-2DE) and crossed immunoelectrophoresis (CIE). In 7 patients both glycoproteins (GPs) IIb and Ilia were absent or reduced to roughly the same extent. In two related patients only a trace of GP Ilb-IIIa complex was detected in CIE, but HR-2DE revealed a glycopeptide in the position of GP Ilia in an amount comparable to type II thrombasthenia. This GP Ilia-like component was neither recognized normally by anti-GP Ilb-IIIa antibodies nor labeled by surface iodination. In unreduced-reduced two-dimensional gel electrophoresis two components were observed in the region of GP Ilia. The assumption of a structural variant of GP Ilia in the two related patients is discussed.

Download Full-text

Specific Binding of Platelet Proteins to Immobilized Fibrin Monomers

Thrombosis and Haemostasis ◽

10.1055/s-0038-1657078 ◽

1979 ◽

Vol 42 (05) ◽

pp. 1686-1689

Author(s):

Czeslaw S Cierniewski ◽

Tadeusz Krajewski ◽

Jolanta Waczyńska-Niewiarowska

Keyword(s):

Specific Binding ◽

Fibrin Monomers ◽

Platelet Proteins

Download Full-text

Human megakaryocytes. II. Expression of platelet proteins in early marrow megakaryocytes.

Journal of Experimental Medicine ◽

10.1084/jem.154.1.88 ◽

1981 ◽

Vol 154 (1) ◽

pp. 88-100 ◽

Cited By ~ 117

Author(s):

E M Rabellino ◽

R B Levene ◽

L L Leung ◽

R L Nachman

Keyword(s):

Factor Viii ◽

Fc Receptors ◽

Platelet Factor 4 ◽

Platelet Factor ◽

Ia Antigen ◽

Plasma Factor ◽

Factor Viii Antigen ◽

Megakaryocyte Differentiation ◽

Marrow Cells ◽

Platelet Proteins

Analysis of various platelet proteins by immunofluorescence demonstrated that platelet glycoproteins Ib, IIb, and IIIa, as well as plasma factor VIII antigen (factor VIII:AGN), platelet factor 4, and fibronectin are present in the vast majority of morphologically recognizable megakaryocytes. In addition, a small number of lymphoid-like mononuclear marrow cells, representing approximately 1.4--2.9/10(4) marrow cells, was found to express the same platelet proteins. This population of early marrow megakaryocytes is analogous to small acetylcholinesterase-positive rat and mouse marrow cells. Fc receptors for IgG were expressed in all megakaryocytes and megakaryocyte precursors, whereas the Ia antigen was detected only on a proportion of mature megakaryocytes and not on only early or precursor megakaryocytes. Platelet glycoproteins Ib, IIb, and IIIa, as well as factor VIII:AGN, and platelet factor 4 were established as distinct markers for marrow megakaryocytes and may be helpful for identifying megakaryocytic cells as well as for monitoring events of megakaryocyte differentiation.

Download Full-text

The pool of fatty acids covalently bound to platelet proteins by thioester linkages can be altered by exogenously supplied fatty acids

Lipids ◽

10.1007/bf02562334 ◽

1999 ◽

Vol 34 (S1) ◽

pp. S331-S337 ◽

Cited By ~ 25

Author(s):

Laszlo Muszbek ◽

Gizella Haramura ◽

Joanne E. Cluette-Brown ◽

Elizabeth M. Van Cott ◽

Michael Laposata

Keyword(s):

Fatty Acids ◽

Platelet Proteins ◽

Covalently Bound

Download Full-text

Radioimmunoassay of Platelet Proteins

Radioimmunoassay in Basic and Clinical Pharmacology - Handbook of Experimental Pharmacology ◽

10.1007/978-3-642-71809-0_21 ◽

1987 ◽

pp. 517-541 ◽

Cited By ~ 1

Author(s):

D. S. Pepper

Keyword(s):

Platelet Proteins

Download Full-text

Cysteine sulfenylation by CD36 signaling promotes arterial thrombosis in dyslipidemia

Blood Advances ◽

10.1182/bloodadvances.2020001609 ◽

2020 ◽

Vol 4 (18) ◽

pp. 4494-4507 ◽

Cited By ~ 1

Author(s):

Moua Yang ◽

Wei Li ◽

Calvin Harberg ◽

Wenjing Chen ◽

Hong Yue ◽

...

Keyword(s):

Reactive Oxygen Species ◽

Hydrogen Peroxide ◽

Platelet Aggregation ◽

Platelet Activation ◽

Arterial Thrombosis ◽

Reactive Oxygen ◽

Src Family Kinases ◽

Oxygen Species ◽

Carbon Nucleophiles ◽

Platelet Proteins

Abstract Arterial thrombosis in the setting of dyslipidemia promotes clinically significant events, including myocardial infarction and stroke. Oxidized lipids in low-density lipoproteins (oxLDL) are a risk factor for athero-thrombosis and are recognized by platelet scavenger receptor CD36. oxLDL binding to CD36 promotes platelet activation and thrombosis by promoting generation of reactive oxygen species. The downstream signaling events initiated by reactive oxygen species in this setting are poorly understood. In this study, we report that CD36 signaling promotes hydrogen peroxide flux in platelets. Using carbon nucleophiles that selectively and covalently modify cysteine sulfenic acids, we found that hydrogen peroxide generated through CD36 signaling promotes cysteine sulfenylation of platelet proteins. Specifically, cysteines were sulfenylated on Src family kinases, which are signaling transducers that are recruited to CD36 upon recognition of its ligands. Cysteine sulfenylation promoted activation of Src family kinases and was prevented by using a blocking antibody to CD36 or by enzymatic degradation of hydrogen peroxide. CD36-mediated platelet aggregation and procoagulant phosphatidylserine externalization were inhibited in a concentration-dependent manner by a panel of sulfenic acid–selective carbon nucleophiles. At the same concentrations, these probes did not inhibit platelet aggregation induced by the purinergic receptor agonist adenosine diphosphate or the collagen receptor glycoprotein VI agonist collagen-related peptide. Selective modification of cysteine sulfenylation in vivo with a benzothiazine-based nucleophile rescued the enhanced arterial thrombosis seen in dyslipidemic mice back to control levels. These findings suggest that CD36 signaling generates hydrogen peroxide to oxidize cysteines within platelet proteins, including Src family kinases, and lowers the threshold for platelet activation in dyslipidemia.

Download Full-text